Anti-IL-27 antibodies and uses thereof

Abstract

The present disclosure relates to anti-IL-27 antibodies and antigen-binding portions thereof. The present disclosure also relates to methods for treating or ameliorating one or more symptoms of a disease, such as cancer, by administering an antibody or antigen-binding portion thereof at a dose of at least about 0.003 mg / kg to at least about 20 mg / kg. The present disclosure generally relates to compositions and methods for modulating IL-27 signaling. More specifically, the present disclosure relates to immunogenic compositions (e.g., antibodies, antibody fragments, and the like) that bind to IL-27 and modulate IL-27 signaling.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. US63 / 185,989, filed May 7, 2021, US63 / 203,688, filed July 28, 2021, and US63 / 277,035, filed November 8, 2021, each of which is incorporated by reference in its entirety herein.

[0002] References to sequence listings submitted electronically via EFS-WEB The contents of the electronically submitted sequence listing (Name: 4416_013PC03_Seqlisting_ST25.txt; Size: 156,680 bytes; and Creation Date: May 6, 2022) are incorporated by reference in their entirety into this specification.

[0003] The present disclosure relates generally to compositions and methods for modulating IL-27 signaling. More particularly, the present disclosure relates to immunogenic compositions (e.g., antibodies, antibody fragments, and the like) that bind to IL-27 and modulate IL-27 signaling. [Background technology]

[0004] In recent years, increasing evidence suggests that the immune system acts as an important barrier against tumor formation and progression. The principle that naturally occurring T cells with antitumor ability or activity exist in cancer patients has rationalized the development of immunotherapy approaches in oncology. Immune cells (such as T cells, macrophages, and natural killer cells) can exhibit antitumor activity and effectively control the appearance 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 avoid immune attacks through various immunoregulatory mechanisms, resulting in failure to control tumor appearance and progression (Motz & Coukos, (2013) Immunity 39(1):61-730). Indeed, an emerging hallmark of cancer is the exploitation of these immunoregulatory mechanisms and the subversion of antitumor immune responses, resulting in tumor evasion and escape from immunological killing (Hanahan and Weinberg (2011) Cell 144(5):646-674).

[0005] IL-27 is a heterodimeric cytokine 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 and mediates signaling through a heterodimeric receptor composed of IL-27Rα (WSX1) and gp130 chains, which preferentially mediate signaling through STAT1 and STAT3. Early reports characterized IL-27 as an immune-promoting 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 shown that IL-27 exhibits complex immunomodulatory functions, resulting in either pro- or anti-inflammatory effects, depending on the biological context and experimental model used. IL-27 drives the expression of different immunomodulatory molecules in human cancer cells, which may support local perturbation of immune responses in vivo (Fabbi et al., (2017) Mediators Inflamm 3958069. Published online February 1, 2017. doi:10.1155 / 2017 / 3958069, and references contained therein). Despite the significant advances that have been made in the treatment and management of cancer, there remains a continuing need for new and effective therapies for the treatment and management of cancer. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Chen & Mellman,(2013)Immunity 39(1):1-10 [Non-Patent Document 2] Motz & Coukos,(2013)Immunity 39(1):61-730 [Non-Patent Document 3] Hanahan and Weinberg (2011) Cell 144(5):646-674 [Non-Patent Document 4] Fabbi et al.,(2017)Mediators Inflamm 3958069. Published online February 1, 2017. doi:10.1155 / 2017 / 3958069 Summary of the Invention

[0007] Some aspects of the disclosure relate to a method of stimulating an immune response in a subject, comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27, wherein the antibody or antigen-binding portion thereof specifically binds to an epitope that includes one or more amino acids of (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142-164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii), and wherein 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.

[0008] Some aspects of the present disclosure relate to a method of treating cancer in a subject in need of treatment comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27, wherein the antibody or antigen-binding portion thereof specifically binds to an epitope that includes one or more amino acids of (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142-164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii), and wherein 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.

[0009] In some embodiments, the antibody or antigen-binding portion thereof is administered at least about 0.003 mg / kg, at least about 0.006 mg / kg, at least about 0.009 mg / kg, at least about 0.03 mg / kg, at least about 0.06 mg / kg, at least about 0.09 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, at least about 10 mg / kg, at least about 12 mg / kg, at least about 14 mg / kg, at least about 16 mg / kg, at least about 18 mg / kg, at least about 19 mg / kg, at least about 20 mg / kg, at least about 21 mg / kg, at least about 22 mg / kg, at least about 23 mg / kg, at least about 24 mg / kg, at least about 25 mg / kg, at least about 26 mg / kg, at least about 27 mg / kg, at least about 28 mg / kg, at least about 29 mg / kg, at least about 30 mg / kg, at least about 31 mg / kg, at least about 32 mg / kg, at least about 33 mg / kg, at least about 34 mg / kg, at least about 35 mg / kg, at least about 36 mg / kg, at least about 37 mg / kg, at least about 38 mg / kg, at least about 39 mg / kg, at least about 40 mg / kg, at least about 41 mg / kg, at least about 42 mg / kg, at least about 43 mg / kg, at least about 44 and / or 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, at least about 10 mg / kg, 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.

[0010] In some embodiments, the antibody or antigen-binding portion thereof is administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 6 weeks, about once every 8 weeks, or about once every 12 weeks.

[0011] In some embodiments, the antibody or antigen-binding portion thereof is administered about once every four weeks. 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 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 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 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 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 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 20 mg / kg about once every four weeks.

[0012] In some embodiments, the antibody or antigen binding portion thereof inhibits or reduces IL-27-dependent STAT1 and / or STAT3 phosphorylation in a cell of the subject. 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 expression of PD-L1 in a cell of the subject. In some embodiments, the antibody or antigen binding portion thereof induces or promotes PD-1-mediated secretion of one or more cytokines from a cell of the subject. In some embodiments, the anti-IL-27 antibody alters expression of TIM-3 in the cell. In some embodiments, the cell is a tumor cell or an immune cell.

[0013] 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, Phe153, 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 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).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).

[0014] 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 (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.

[0015] 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 or 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 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.

[0016] In some embodiments, the antibody or antigen-binding portion thereof comprises: (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.

[0017] In some embodiments, the antibody or antigen-binding portion thereof comprises: (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.

[0018] 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.

[0019] 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.

[0020] 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 an amino acid sequence set forth in SEQ ID NO: 125. 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 heavy chain variable region comprising an 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.

[0021] 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.

[0022] 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), 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.

[0023] 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 the antibody or its antigen-binding portion, after the antibody or its antigen-binding portion, or simultaneously with the antibody or its antigen-binding portion. In some embodiments, the additional therapeutic agent comprises chemotherapy, targeted anti-cancer therapy, oncolytic drug, cytotoxic agent, immune-based therapy, cytokine, surgical procedure, radiation treatment, activator of costimulatory molecule, inhibitor of inhibitory molecule, vaccine, cellular immunotherapy, biologic agent, or combination thereof. In some embodiments, the additional therapeutic agent comprises PD-1 antagonist, PD-L1 inhibitor, TIM-3 inhibitor, LAG-3 inhibitor, TIGIT inhibitor, CD112R inhibitor, TAM inhibitor, STING agonist, 4-1BB agonist, or combination thereof.

[0024] In some embodiments, the additional therapeutic agent comprises a PD-1 antagonist. In 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. 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 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, 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.In some embodiments, the additional therapeutic agent is a TIM-3 inhibitor. In some embodiments, the TIM-3 inhibitor is MGB453 or TSR-022. 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. 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).

[0025] 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 EBI3, IL-27, TNFα, MIP-1α (CCL3), IFNγ, IL-10, IL-6, and any combination thereof, and the increased expression of the one or more biomarkers is compared 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 EBI3, and the increased expression of EBI3 is compared to the expression of EBI3 prior to administration.

[0026] 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, and the increased expression of the one or more biomarkers is compared 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), and the increased expression of eotaxin-1 (CCL11) is compared to the expression of eotaxin-1 (CCL11) prior to administration.

[0027] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits an increase in circulating concentrations of IFNγ compared to the circulating concentrations of IFNγ prior to administration. [Brief description of the drawings]

[0028] [Figure 1A] Schematic diagram of a Phase 1 dose escalation study of an anti-IL-27 antibody. ccRCC=clear cell renal cell carcinoma; CR=complete response; HCC=hepatocellular carcinoma; N=number; PR=partial response; pts=patients, RP2D=recommended Phase 2 dose; 2L=secondary. Dose level 1=0.003mg / kg; dose level 2=0.03mg / kg; dose level 3=0.1mg / kg; dose level 4=0.3mg / kg; dose level 5=1.0mg / kg; dose level 6=3.0mg / kg; dose level 7=10.0mg / kg; dose level 8=20.0mg / kg. [Figure 1B] FIG. 1 is a schematic diagram of a Phase 1 dose escalation study of an anti-IL-27 antibody. [Figure 1C] FIG. 1 is a schematic diagram of a Phase 1 dose escalation study of an anti-IL-27 antibody. [Diagram 2] 1 is a swimmer plot showing study duration and RECIST responses grouped by starting dose. [Diagram 3] FIG. 13 is a waterfall plot showing best percent change from baseline in target lesions. [Figure 4]1 is a graphic representation of the pharmacokinetics of anti-IL-27 Ab1 administered at doses of 0.03, 0.1, 0.3, 1, 3, and 10 mg / kg. [Figure 5A] FIG. 1 is a graphical representation of T cells gated using anti-CD3 antibody. [Figure 5B] This is a graph showing the analysis of T cells using anti-pSTAT1 Y701 antibody and comparison with the results before administration. [Figure 5C] 13 is a bar graph showing pSTAT1 inhibition before and after dosing on day 1 of cycle 2 with 0.1 mg / kg anti-IL-27 Ab1. [Figure 5D] 13 is a bar graph showing pSTAT1 inhibition before and after 1 mg / kg anti-IL-27 Ab1 dosing on day 1 of cycle 2. [Figure 6] A-F are images of target lesions (target lesion 1, A-C; target lesion 2, C-F; arrows) in a 64-year-old patient with squamous non-small cell lung cancer with mediastinal lymph node, lung, and pleural metastases enrolled at 10 mg / kg. A and D are images at baseline, B and E are images at week 8, and C and F are images at week 12. [Figure 7] Swimmer plot showing study duration and RECIST response grouped by starting dose for 29 patients enrolled in the dose escalation study. Median study duration was 9 weeks (range 1-71 weeks). [Figure 8A] Changes in target lesions over time. Waterfall plot showing best percent change from baseline in target lesions (n=27). [Figure 8B] Figure 1 shows the change in target lesions over time. Spider plot showing the change in target lesions over time from baseline. [Figure 9A] Graphical representation of dose escalation response of anti-IL-27 Ab1 monotherapy.Waterfall plot showing best percent change in total target lesions. [Figure 9B] Graphical representation of dose escalation response of anti-IL-27 Ab1 monotherapy.Spider plot showing lesions over time. [Figure 10A]1 is a graphical representation of anti-IL-27 Ab1 pharmacokinetic profile. Cycle 1 anti-IL-27 Ab1 PK by dose regimen across tumor types. [Figure 10B] 1 is a graphical representation of the pharmacokinetic profile of anti-IL-27 Ab1. PK of anti-IL-27 Ab1 when administered once every 4 weeks as monotherapy in the dose escalation phase of the study (mixed solid tumors) and in expanding HCC and ccRCC. [Figure 11A] Graphical representation of dose escalation response of anti-IL-27 Ab1 to ccRCC monotherapy. Waterfall plot showing best percent change in total target lesions. [Figure 11B] Graphical representation of dose escalation response of anti-IL-27 Ab1 to ccRCC monotherapy. Spider plot showing lesion evolution over time. [Figure 12A] 1 is a graphical representation showing the efficacy of anti-IL-27 Ab1 as monotherapy for HCC. Waterfall plot showing best percent change in total target lesions. [Figure 12B] 1 is a graphical representation showing the efficacy of anti-IL-27 Ab1 as monotherapy for HCC. 2 is a spider plot showing lesion progression over time. [Figure 13A] 1 is a graphical representation of IL-27-dependent pSTAT1 inhibition in T cells from patient blood following administration of 0.1 mg / kg, 1.0 mg / kg, and 3.0 mg / kg of anti-IL-27 antibody (y-axis) and the corresponding serum levels of anti-IL-27 antibody following administration of anti-IL-27 antibody (x-axis). The dotted vertical line represents the serum concentration of anti-IL-27 antibody that results in 90% (IC90) inhibition of IL-27-dependent pSTAT1 (0.7 μg / ml). [Figure 13B] Pharmacokinetic analysis of anti-IL-27 antibody in the serum of a subject after repeated dosing of 0.1 mg / kg of anti-IL-27 antibody once every 28 days is shown. The serum level of anti-IL-27 antibody required to achieve IC90 for IL-27-dependent inhibition of pSTAT1 in blood-derived T cells of the subject is shown by the horizontal dotted line. [Figure 13C]1 shows a pharmacokinetic analysis of anti-IL-27 antibody in the serum of a subject following repeated dosing of 1.0 mg / kg of anti-IL-27 antibody once every 28 days. The serum level of anti-IL-27 antibody required to achieve an IC90 for IL-27-dependent inhibition of pSTAT1 in blood-derived T cells of the subject is indicated by the horizontal dotted line. [Figure 13D] 1 shows a pharmacokinetic analysis of anti-IL-27 antibody in the serum of a subject after repeated dosing of 3.0 mg / kg of anti-IL-27 antibody once every 28 days. The serum level of anti-IL-27 antibody required to achieve an IC90 for IL-27-dependent inhibition of pSTAT1 in blood-derived T cells of the subject is indicated by the horizontal dotted line. [Figure 14] A is a graphical representation of the fold change in eotaxin-1 versus baseline at 6 hours post-C1D1 administration in patients who showed progressive disease (PD), stable disease (SD), or partial response (PR) following administration of various doses of anti-IL-27 Ab1. B is a graphical representation of the longitudinal analysis of the fold change in eotaxin-1 versus baseline across time points C1D1 (pre-dose and 6 hours post-dose), C1D8, C2D1 (pre-dose and 6 hours post-dose), and C3D1. Data from patients who showed partial response are labeled "PR." Each data set represents one patient. [Figure 15] 1 shows fold change in IL-27 levels versus baseline at different visits, times, and dosage cohorts for anti-IL-27 Ab1 monotherapy patients. [Figure 16] 1 shows the fold change in circulating IFNγ levels versus baseline at various visit and time cohorts for 10 mg / kg anti-IL-27 Ab1 monotherapy patients. [Figure 17A] Graphical representation of fold change in expression of TARC (CCL17) versus baseline in samples from patients treated with anti-IL-27 Ab1. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor shrinkage are labeled. [Figure 17B]Graphical representation of fold change in expression of VEGF-A versus baseline in samples from patients treated with anti-IL-27 Ab1. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor shrinkage are labeled. [Figure 17C] Graphical representation of fold change in expression of IL-7 versus baseline in samples from patients treated with anti-IL-27 Ab1. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor shrinkage are labeled. [Figure 17D] Graphical representation of fold change in expression of IL-8 versus baseline in samples from patients treated with anti-IL-27 Ab1. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor shrinkage are labeled. [Figure 17E] Graphical representation of fold change in expression of MCP-1 versus baseline in samples from patients treated with anti-IL-27 Ab1. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor shrinkage are labeled. [Figure 17F] 1 is a graphical representation of the fold change in expression of MCP-4 versus baseline in samples from patients treated with anti-IL-27 Ab1. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor regression are labeled. [Figure 17G] 17A-17F are waterfall plots showing fold change from baseline in target lesions of patients characterized in Figures 17A-17F. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) who showed tumor shrinkage are labeled. [Figure 18A]FIG. 1 is a graphical representation of longitudinal analysis of IL-7, TARC (CCL17), and VEGF-A in samples from patients treated with anti-IL-27 Ab1, as described above. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) but tumor shrinkage are labeled. [Figure 18B] FIG. 1 is a graphical representation of longitudinal analysis of IL-8, MCP-1, and MCP-4 in samples from patients treated with anti-IL-27 Ab1, as described above. Data corresponding to patients with confirmed partial response (PR; 902-002) and patients with stable disease (SD; 901-008) but tumor shrinkage are labeled. [Figure 19] FIG. 11 is a scatter plot showing IL-27-induced changes in gene expression from two individuals. [Figure 20] AC are scatter plots showing gene expression changes from two individuals following exposure to IL-27 heterodimer (A), EBI3 alone (B), or IL-35 (C). [Figure 21] A-B are volcano plots representing gene set enrichment analysis of the IL-27 gene signature from CD4+ T cells. In A, enrichment of mRNA signatures related to interferon signaling is highlighted (grey). In B, characteristic IFNα signature genes are highlighted (grey). [Figure 22A] Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml), showing clustering of different immune cell types. [Figure 22B] Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml). Volcano plot showing IL-27-mediated gene expression changes identified in the total PBMC population, including many interferon-stimulated genes. [Figure 22C]Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml).Volcano plot showing down- and up-regulation of IL-27 signature genes in immune cell subpopulations of NK cells. [Figure 22D] Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml).Volcano plot showing down- and up-regulation of IL-27 signature genes in immune cell subpopulations of CD4+ T cells. [Figure 22E] Graphical representation of single cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml).Volcano plot showing down- and up-regulation of IL-27 signature genes in immune cell subpopulations of B cells. [Figure 22F] Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml).Volcano plot showing down- and up-regulation of IL-27 signature genes in monocytic immune cell subpopulations. [Figure 22G] Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml).Volcano plot showing down- and up-regulation of IL-27 signature genes in immune cell subpopulations of CD8+ T cells. [Fig. 22H]Graphical representation of single-cell RNA sequencing analysis of PBMCs stimulated in vitro with anti-CD3 (0.25 μg / ml) in the presence or absence of rhIL-27 (100 ng / ml).Volcano plot showing down- and up-regulation of IL-27 signature genes in immune cell subpopulations of Treg cells. [Figure 23A] FIG. 1 is a bar graph showing the assessment of IL-17A in culture supernatants of pooled PBMCs activated for 4 days in the presence of various cytokines (100 ng / ml; x-axis) in the presence of anti-CD3 (0.25 μg / ml) and anti-PD-1 (1 μg / ml). [Figure 23B] FIG. 1 is a bar graph showing the assessment of IFN-γ in culture supernatants of pooled PBMCs activated for 4 days in the presence of various cytokines (100 ng / ml; x-axis) in the presence of anti-CD3 (0.25 μg / ml) and anti-PD-1 (1 μg / ml). [Figure 24A] Clustering of different immune cell types based on single-cell RNA-seq analysis of IL27 expression. [Figure 24B] FIG. 11 is a scatter plot showing that the MF2 macrophage gene signature associated with disease progression contains several interferon-stimulated genes and is highly enriched in IL27-positive compared to IL27-negative macrophages. [Figure 24C] 1 is a graphical representation showing that IL27 expression is increased in macrophages from patients with disease progression. [Figure 24D] FIG. 1 is a graphical representation showing that IL27 expression is increased in macrophages from metastatic and primary tumors compared to normal tissue. [Figure 24E] 1 is a graphical representation showing that IL27 expression is increased in macrophages from patients with stage IV disease. [Figure 24F] Volcano plot showing MF2 signature genes (grey) of monocyte-derived macrophages stimulated with IL-27 in vitro. [Figure 24G]Immunohistochemistry images for IL-27 on tissue microarrays showing positive expression in macrophages in the TME of lung adenocarcinoma (AdenoCa). [Fig. 24H] Immunohistochemistry images for IL-27 on tissue microarrays showing positive expression in macrophages in the TME of squamous cell carcinoma (SCC). [Figure 25A] Figure 1 shows the clustering of different cell types in the tumor microenvironment based on single-cell RNA-seq analysis of IL27 expression. [Figure 25B] Violin plot of IL27RA expression in different cell populations compared to tumor cells. [Figure 25C] FIG. 13 is a violin plot of IL27RA expression in tumor cells from patients with progressive disease compared to tumor cells from patients with residual disease or treatment-naive patients. [Figure 26A] 1 is a bar graph showing expression of IL27RA mRNA transcripts across the Cancer Cell Line Encyclopedia (CCLE) for various lung cancer cell lines, including NCI-H2228. [Figure 26B] 1 is a graphic representation showing pSTAT1 levels in NCI-H2228 lung cancer cells after IL-27 stimulation. [Figure 26C] 1 is a graphic representation showing PDL1 expression in NCI-H2228 lung cancer cells after IL-27 stimulation. [Figure 26D] Graphical representation of microarray profiling of NCI-H228 cells cultured for 48 hours in the presence or absence of IL-27. Several interferon-responsive genes are flagged. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] Some embodiments of the present disclosure relate to a method of stimulating an immune response in a subject, comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27 or an antigen-binding portion thereof. Some embodiments of the present disclosure relate to a method of treating cancer in a subject in need of such treatment, comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27. 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 that includes one or more amino acids of (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142-164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii).

[0030] I. Definition Terms used in the claims and specification, unless otherwise specified, are defined as set forth below.

[0031] 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 dictates otherwise.

[0032] 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 one of ordinary skill in the art, "about" will mean up to plus or minus 10% of the particular value, taking into account the context in which it is used.

[0033] As used herein, the term "agonist" refers to any molecule that partially or fully enhances, induces, increases, 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 by a negative control under comparable conditions. Methods for identifying agonists suitable for use in the methods of the present disclosure are also disclosed herein. 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 thus indicate the ability of the agonist to enhance, 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 an agonist to activate or enhance 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 usually measured by its EC 50 The EC value is defined as the concentration required to activate 50% of the agonist response. 50The lower the value, the more potent the agonist is and the lower the concentration required to activate a maximal biological response.

[0034] As used herein, the term "alanine scanning" refers to a technique used to determine the contribution of a specific wild-type residue to the stability or function(s) (e.g., binding affinity) of a given protein or polypeptide. The technique involves substituting alanine residues for wild-type residues in a polypeptide, followed by assessing the stability or function(s) (e.g., binding affinity) of the alanine-substituted derivative or mutant polypeptide and comparing it to the wild-type polypeptide. Techniques for substituting alanine for wild-type residues in a polypeptide are known in the art.

[0035] The term "amelioration" refers to any therapeutically beneficial outcome in the treatment of a disease state (e.g., cancer), including prevention, reduction in severity or progression, remission, or cure.

[0036] As used herein, the term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that have been subsequently modified (e.g., hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine). Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid (i.e., a hydrogen, a carboxyl group, an amino group, and a carbon bonded to an R group) (e.g., homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium). Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refer to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that function in a manner similar to a naturally occurring amino acid.

[0037] 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.

[0038] As used herein, an "amino acid substitution" refers to the replacement of at least one existing amino acid residue in a given amino acid sequence (the amino acid sequence of the starting polypeptide) with a second, different, "replacement" amino acid residue. An "amino acid insertion" refers to the incorporation of at least one additional amino acid into a given amino acid sequence. An insertion usually consists of the insertion of one or two amino acid residues, although larger "peptide insertions" can also be made (e.g., insertions of about 3 to about 5, or even up to about 10, 15, or 20 amino acid residues). The inserted residue(s) can be naturally occurring or non-naturally occurring, as disclosed above. An "amino acid deletion" refers to the removal of at least one amino acid residue from a given amino acid sequence.

[0039] As used herein, the term "amount" or "level" is used in the broadest sense and refers to the amount, concentration, or abundance of a substance (e.g., a metabolite, a small molecule, a protein, an mRNA, a marker). When referring to a metabolite or a small molecule (e.g., a drug), the terms "amount," "level," and "concentration" are generally used interchangeably and generally refer to a detectable amount in a biological sample. "Elevated level" or "increased level" refers to an increase in the amount, concentration, or abundance of a substance in a sample compared to a control sample (such as from an individual(s) not afflicted with a disease or disorder (e.g., cancer), or an internal control). In some embodiments, an elevated level of a substance (e.g., a drug) in a sample refers to an increase in the amount of a 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% compared to the amount of the substance in a control sample, as determined by techniques known in the art (e.g., HPLC). A "reduced level" refers to a decrease in the amount, concentration, or abundance of a substance (e.g., a drug) in an individual compared to a control (such as from an individual(s) not afflicted with a disease or disorder (e.g., cancer) or an internal control). In some embodiments, a reduced level refers to little or no detectable amount, concentration, or abundance. In some embodiments, a reduced level of a substance (e.g., a drug) 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% compared to the amount of the substance in a control sample as determined by techniques known in the art (e.g., HPLC).

[0040] When referring to a protein, mRNA, or marker (such as those described herein), the term "level of expression" or "expression level" is generally used interchangeably and generally refers 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 related to the likelihood of a response to an agent (such as those described herein). "Expression" generally refers to the process by which information contained within a gene is converted into a structure (e.g., a protein marker such as PD-L1) that is present and operational in a cell. Thus, as used herein, "expression" can refer to transcription into a polynucleotide, translation into a polypeptide, or, in some cases, modification of a polynucleotide and / or polypeptide (e.g., post-translational modification of a polypeptide). Fragments of a transcribed polynucleotide, a translated polypeptide, or modification of a polynucleotide and / or polypeptide (e.g., post-translational modification of a polypeptide) will also be considered as expressed, regardless of whether they arise from a transcript generated by alternative splicing or a degraded transcript, or from post-translational processing of a polypeptide, for example, by proteolysis. "Expressed genes" include those that are transcribed into polynucleotides as mRNA and then translated into polypeptides, and those that are further transcribed into RNA but not translated into polypeptides (e.g., transfer and ribosomal RNA). "Elevated expression," "elevated expression level," or "elevated level" refers to an increase in expression or an increase in the level of a substance in a sample compared to a control sample (such as an individual(s) not afflicted with a disease or disorder, e.g., cancer, or an internal control).In some embodiments, elevated expression of a substance (e.g., a protein marker such as PD-L1) in a sample refers to an increase in the amount of a 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% compared 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 a decrease in the level of a substance (e.g., a protein marker) in an individual compared to a control (such as an individual(s) 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, reduced 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% compared to the amount of the substance in a control sample as determined by techniques known in the art (e.g., FACS).

[0041] As used herein, the term "angiogenesis" or "angiogenesis" refers to the process by which new blood vessels develop 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, either from pre-existing blood vessels or from circulating endothelial stem cells (Takahashi et al., (1995) Nat. Med. 5:434-438; Isner et al., (1999) J. Clin. Invest. 103:1231-1236), become activated to migrate, proliferate, and differentiate into luminal structures to form new blood vessels in response to growth factor or hormonal cues, or hypoxic or ischemic conditions. During ischemia (such as occurs in cancer), the need for increased oxygen supply and nutrient delivery apparently induces the secretion of angiogenic factors by the affected tissue, and these factors stimulate the formation of new blood vessels. Several additional terms relate to angiogenesis.

[0042] The term "antagonist" as used herein refers to an inhibitor of a target molecule, and may be used herein synonymously with the term "inhibitor". As used herein, the term "antagonist" refers to any molecule that partially or fully blocks, inhibits, or neutralizes the biological activity of a native polypeptide disclosed herein. Suitable antagonist molecules 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 antagonist 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 by a negative control under comparable conditions. Methods for identifying antagonists suitable for use in the disclosed methods are also disclosed herein. For example, these methods include, but are not limited to, binding assays such as enzyme-linked immunosorbent assays (ELISAs), the ForteBio® system, 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 antagonist to bind to a polypeptide of interest (e.g., a receptor or ligand), and thus indicate the ability of the antagonist to inhibit, neutralize, or block the activity of the polypeptide. The effectiveness of an antagonist can also be determined using functional assays (such as the ability of an antagonist to inhibit the function of a polypeptide).For example, a functional assay may involve contacting a polypeptide with a candidate antagonist molecule and measuring a detectable change in one or more biological activities normally associated with the polypeptide. The potency of an antagonist is usually measured by its IC. 50 The IC value (the concentration required to inhibit 50% of the agonist response) is used to define the inhibitory effect of an agonist. 50 The lower the value, the more potent the antagonist is and the lower the concentration required to inhibit the maximal biological response.

[0043] 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., with respect to a decrease (or reduction) of human IL-27 activity of at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. Additional examples of IL-27 biological activity and / or downstream pathway(s) mediated by IL-27 signaling, or other IL-27-mediated function are described in additional detail below and elsewhere herein.

[0044] As used herein, the term "anti-IL-27 antagonist antibody" (interchangeably termed "anti-IL-27 antibody") refers to an antibody that specifically binds to IL-27 and inhibits IL-27 biological activity and / or downstream pathway(s) mediated by IL-27 signaling, or other IL-27-mediated function. Anti-IL-27 antagonist antibodies encompass antibodies that block, antagonize, inhibit, inhibit, or reduce the biological activity of IL-27 (e.g., ligand binding, enzymatic activity), including downstream pathways mediated by IL-27 signaling or function (such as receptor binding to and / or 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 to 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 antibodies prevent, block, or inhibit binding of human IL-27 to gp130. In some embodiments, the anti-IL-27 antagonist antibodies prevent, block, or inhibit binding of human IL-27 to IL-27Rα. In some embodiments, the anti-IL-27 antagonist antibodies prevent, block, or inhibit dimerization of IL-27 monomers. In some embodiments, the anti-IL-27 antibodies do not specifically bind to EBI3 monomers. In some embodiments, the anti-IL-27 antibody specifically binds to IL-27p28 monomer. In some embodiments, the anti-IL-27 antibody specifically binds to a non-contiguous epitope that includes p28, but does not bind to EBI3 monomer. In some embodiments, the anti-IL-27 antibody inhibits or reduces phosphorylation of STAT1 and / or STAT3 in a cell. In some embodiments, the anti-IL-27 antibody inhibits or reduces inhibition of CD161 expression in a cell (e.g., ameliorate or alleviate IL-27-mediated inhibition of CD161 expression in a cell).In some embodiments, the anti-IL-27 antibody inhibits or reduces the expression of PD-L1 in a cell. In some embodiments, the anti-IL-27 antibody induces or promotes PD-1-mediated secretion of one or more cytokines from a cell. In some embodiments, the anti-IL-27 antibody alters the expression of TIM-3 in a cell. In some embodiments, the anti-IL-27 antagonist antibody binds to human IL-27 and stimulates or promotes 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 mutant IgG1 heavy chain constant region, or a wild-type or mutant IgG4 heavy chain constant region. Examples of anti-IL-27 antagonist antibodies are provided herein.

[0045] As used herein, the term "antibody" refers to a complete antibody comprising two light chain polypeptides and two heavy chain polypeptides. Complete antibodies include different antibody isotypes including IgM, IgG, IgA, IgD, and IgE antibodies. The term "antibody" includes polyclonal, monoclonal, chimeric or chimeric, humanized, primatized, deimmunized, and fully human antibodies. Antibodies can be made in or derived from any of a variety of species (e.g., mammals such as humans, non-human primates (e.g., orangutans, baboons, or chimpanzees), horses, cows, pigs, sheep, goats, dogs, cats, rabbits, guinea pigs, gerbils, hamsters, rats, and mice). Antibodies can be purified or recombinant. As used herein, the terms "antibody fragment", "antigen-binding fragment", or similar terms refer to a fragment of an antibody that retains the ability to bind to a target antigen (e.g., IL-27) and inhibit the activity of the target antigen. 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 that contains both the heavy and light chain variable regions of the antibody from which the scFv is derived. In addition, intrabodies, minibodies, triabodies, and diabodies are also included within the definition of an antibody and are compatible 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 in their entireties.

[0046] As used herein, the term "antibody fragment" also encompasses, for example, single domain antibodies (such as camelized single domain antibodies). See, e.g., 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 Publication Nos. WO 94 / 04678 and WO 94 / 25591; and U.S. Patent No. 6,005,079, all of which are incorporated herein by reference in their entirety. In some embodiments, the disclosure provides single domain antibodies comprising two VH domains with modifications such that a single domain antibody is formed.

[0047] In some embodiments, the antigen-binding fragment comprises a variable region of a heavy chain polypeptide and a variable region of a light chain polypeptide. In some embodiments, the antigen-binding fragment described herein comprises polypeptides of the light and heavy chain CDRs of the antibody.

[0048] The term "antigen-presenting cell" or "APC" is a cell that presents foreign antigens 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 (such as THP-1), B lymphoblastoid cells (such as C1R.A2, 1518 B-LCL), and monocyte-derived dendritic cells (DCs). Some APCs internalize antigens by phagocytosis or by receptor-mediated endocytosis.

[0049] The term "antigen presentation" refers to the process by which APCs capture antigens and enable their recognition by T cells, for example as components of MHC-I and / or MHC-II complexes.

[0050] 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 result in apoptosis can lead to observable and characteristic morphological changes to cells, including membrane blebbing, cell volume shrinkage, chromosomal DNA condensation and fragmentation, and mRNA decay. A common method for identifying cells, including T cells, undergoing apoptosis is to expose the cells to a fluorophore-conjugated protein (annexin V). Annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine on the outer leaflet of the plasma membrane, which is an early indicator that cells are undergoing the apoptotic process.

[0051] As used herein, the term "B cell" (alternatively "B lymphocyte") refers to a type of white blood cell of the lymphocyte subtype. B cells function in the humoral immune component of the adaptive immune system by secreting antibodies. B cells also present antigens and secrete cytokines. B cells differ from the other two classes 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 against which they will mount an antibody response.

[0052] As used herein, the term "binds to immobilized IL-27" refers to the ability of an antibody of the disclosure to bind to IL-27, e.g., expressed on the surface of a cell or attached to a solid support.

[0053] As used herein, the term "bispecific antibody" 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 a variety of methods, including fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai & Lachmann, (1990) Clin. Exp. Immunol. 79:315-321; Kostelny et al., (1992) J. Immunol. 148:1547-1553.

[0054] Traditionally, recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy / light chain pairs, where the two heavy / light chain pairs have 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 fusion of the heavy chain variable region is preferably with an immunoglobulin heavy chain constant domain, including at least a portion of the hinge, CH2, and CH3 regions. For further details of exemplary currently known methods for generating bispecific antibodies, see, e.g., Suresh et al., (1986) Methods Enzymol. 121:210; PCT Publication No. 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. See, e.g., J. Immunol. 147:60, (1991). 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.

[0055] Various techniques have also been described for producing and isolating bispecific antibody fragments directly from recombinant cell culture. For example, bispecific antibodies have been produced using leucine zippers. See, for example, Kostelny et al. (1992) J Immunol 148(5):1547-1553. The leucine zipper peptides from the 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 utilized for the production of antibody homodimers. The "diabody" technology described by Hollinger et al. (1993) Proc Natl Acad Sci USA 90:6444-6448 provided an alternative mechanism for making bispecific antibody fragments. The fragments contain 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 paired with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by using single-chain Fv (scFv) dimers has also been reported. See, for example, Gruber et al. (1994) J Immunol 152:5368. Alternatively, the antibody can 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.

[0056] 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.

[0057] The present disclosure also encompasses variant forms of multispecific antibodies, such as 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, either directly or via a short linker, followed by a light chain constant domain by recombinant DNA techniques. Similarly, the heavy chain comprises two different heavy chain variable domains (VH) linked in tandem, followed by the constant domain CH1 and Fc region. Methods for generating DVD-Ig molecules from two parent antibodies are further described, for example, in PCT Publication Nos. WO08 / 024188 and WO07 / 024715. In some embodiments, the bispecific antibody is a Fabs-in-Tandem immunoglobulin in which a light chain variable region with a second specificity is fused to a heavy chain variable region of a complete antibody, such as those described in International Patent Application Publication No. WO2015 / 103072.

[0058] 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) fetal 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) other types of antigens or antigen-presenting cells or materials associated with cancer.

[0059] As used herein, the term "cancer-specific immune response" refers to an immune response induced by the presence of tumor, cancer cell, or cancer antigen. In certain embodiments, the response includes the proliferation and division of cancer antigen-specific lymphocytes. In certain embodiments, the 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 and acquired immune systems interact to initiate antigenic responses against tumor, cancer cell, or cancer antigen. In certain embodiments, the cancer-specific immune response is a T cell response.

[0060] The term "carcinoma" is art-recognized and refers to malignant tumors of epithelial or endocrine tissues, including respiratory, gastrointestinal, genitourinary, testicular, breast, prostate, endocrine, and melanoma. The anti-IL-27 antibodies described herein may be used to treat patients with, suspected of having, or who may be at high risk of developing any type of cancer (including renal or melanoma) or any viral disease. Exemplary cancers include those formed from tissues of the cervix, lung, prostate, breast, head and neck, colon, and ovary. The term also encompasses carcinosarcoma, which includes malignant tumors composed of carcinomatous and sarcomatous tissue. "Adenocarcinoma" refers to carcinomas derived from glandular tissue or in which the tumor cells form recognizable glandular structures.

[0061] As used herein, the term "CD112R" refers to a member of the poliovirus receptor-like protein and is a co-inhibitory receptor for human T cells. CD112R is an inhibitory receptor expressed primarily by T cells and NK cells, and competes with the activating receptor CD226 for CD112 binding. The interaction of CD112 with CD112R is of higher affinity than with CD226, thereby effectively modulating CD226-mediated cell activation. Anti-CD112R antagonists that block the interaction with CD112 limit the inhibitory signaling immediately downstream of CD112R, while at the same time promoting greater immune cell activation by increasing CD226 interaction with CD112. As used herein, the term "CD112R inhibitor" refers to an agent that disrupts, blocks, or inhibits the biological function or activity of CD112R.

[0062] As used herein, the term "CD137" (alternatively "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. Crosslinking of CD137 promotes 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 utomirumab (PF-05082566), a fully human IgG2 monoclonal antibody that targets 4-1BB to stimulate T cells.

[0063] As used herein, the term "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 expression of CD161 has been associated with infiltration of T cells into the tumor microenvironment for a number of 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.

[0064] 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 that includes 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 and IL-27 subunit α and IL-27A). IL-27 is preferentially 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, many 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). Although IL-27 was first described as a factor that enhances the initiation of Th1 responses, IL-27 was later found to perform a major T cell suppressive function by limiting Th1 responses, inhibiting differentiation of Th2 and Th17 cells, and regulating 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 immunoregulator, IL-27 also regulates angiogenesis, hematopoiesis, and osteoclastogenesis.

[0065] IL-27 signals through a heterodimeric type I cytokine receptor (IL-27 receptor or IL-27R) that includes a first subunit known as WSX1 (also known as IL-27 receptor subunit alpha, IL-27RA, T cell cytokine receptor type 1 (TCCR), and cytokine receptor-like 1 (CRL1)), and a second subunit known as gp130 (also known as interleukin-6 signaling factor (IL6ST), interleukin-6 receptor subunit beta (IL-6RB), and oncostatin M receptor). gp130 is also the receptor subunit for the IL-6 family of 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 and p38 MAPK pathways.

[0066] EBI3 also appears 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).

[0067] 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-mgqtagdlgwrlsllllplllvqagvwgfprppgrpqlslqelrreftvslhlarkllsevrgqahrfaeshlpgvnlyllplgeqlpdvsltfqawrrlsdperlcfisttlqpfhallgglgtqgrwtnmermqlwamrldlrdlqrhlrfqvlaagfnlpeeeeeeeeeeeeerkgllpgalgsalqgpaqvswpqllstyrllhslelvlsravrellllskaghsvwplgfptlspqp-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).

[0068] As used herein, the term "compete," 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 assays; cross-blocking assays) where a test antigen binding protein (e.g., a test antibody) inhibits (e.g., reduces or blocks) specific binding of a reference antigen binding protein (e.g., a reference antibody) to a common antigen (e.g., IL-27 or a fragment thereof).

[0069] A polypeptide sequence or amino acid sequence "derived from" a specified polypeptide or protein refers to the origin of the polypeptide. Preferably, a polypeptide sequence 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 at least 10-20 amino acids, preferably at least 20-30 amino acids, more preferably at least 30-50 amino acids, or is otherwise identifiable by the skilled artisan as having its origin in the sequence. A polypeptide derived from another peptide may have one or more mutations (e.g., one or more amino acid residues replaced by another amino acid residue or have one or more amino acid residues inserted or deleted) compared to the starting polypeptide.

[0070] Polypeptides may include non-naturally occurring amino acid sequences. Such variants necessarily have less than 100% sequence identity or similarity with the starting molecule. In certain embodiments, variants will have an amino acid sequence identity or similarity of about 75%-100% or less, more preferably about 80%-100% or less, more preferably about 85%-100% or less, more preferably about 90%-100% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%), and most preferably about 95%-100% to the amino acid sequence of the starting polypeptide, for example over the length of the variant molecule.

[0071] In certain embodiments, the antibodies of the present disclosure are encoded by nucleotide sequences that 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 generation for, e.g., passive immunization, PCR, generation of primers and probes, and the like.

[0072] It will also be understood by those skilled in the art that antibodies suitable for use in the methods disclosed herein may be altered to vary in sequence from the naturally occurring or native sequences 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 at "non-essential" amino acid residues may be made. Mutations may be introduced by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis.

[0073] Antibodies suitable for use in the methods disclosed herein may include conservative amino acid substitutions at one or more amino acid residues (e.g., 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 having 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), beta-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 by another amino acid residue from the same side chain family. In certain embodiments, a string of amino acids can be replaced by a structurally similar string that differs in the order and / or composition of side chain family members. Alternatively, in certain embodiments, mutations can be introduced randomly, such as by saturation mutagenesis, along all or a portion of a coding sequence, and the resulting mutants can be incorporated into a binding polypeptide of the present disclosure and screened for their ability to bind to a desired target.

[0074] As used herein, the term "cross-presentation" of an antigen refers to the presentation of a foreign protein antigen to a T cell via MHC class I and MHC class II molecules on an APC.

[0075] As used herein, the term "cross-react" refers to the ability of an antibody of the present disclosure to bind to IL-27 from a different species. For example, an antibody of the present disclosure that binds to human IL-27 may also bind to IL-27 from 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 binding to or otherwise functional interaction with cells that physiologically express IL-27. Methods for determining cross-reactivity include standard binding assays described herein, such as Biacore™ surface plasmon resonance (SPR) analysis using a Biacore™ 2000 SPR instrument (Biacore AB and Uppsala, Sweden), or flow cytometry techniques.

[0076] 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.

[0077] As used herein, the term "dendritic cell" or "DC" refers to a type of antigen-presenting cell that is a bone marrow (BM)-derived white blood cell and is the most potent type of antigen-presenting cell. DC capture and process antigens, converting proteins into peptides that are presented on major histocompatibility complex (MHC) molecules that are recognized by T cells. DC are heterogeneous (e.g., myeloid DC and plasmacytoid DC). Although all DC are capable of uptake, processing, and presenting antigens to naive T cells, subtypes of DC have distinct markers and differ in their location, migration route, detailed immunological functions, and dependence on infection or inflammatory stimuli for their generation. During the development of adaptive immune responses, DC phenotype and function play a role in initiating tolerance, memory, and polarized T-helper 1 (Th1), Th2, and Th17 differentiation.

[0078] As used herein, the term "dendritic cell activation" refers to the transition from immature dendritic cells to mature dendritic cells, where activated dendritic cells encompass mature dendritic cells and dendritic cells in the process of transition, and the expression of CD80 and CD86, which induce costimulatory signals, is elevated by activation stimuli. Mature human dendritic cells are cells positive for the expression of 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, for example, CD80 and CD86. Immature dendritic cells are weakly positive, preferably negative, for these markers, while mature dendritic cells are positive. Differentiation of mature dendritic cells is routinely performed by those skilled in the art, and the respective markers described above and methods for measuring their expression are also well known to those skilled in the art.

[0079] As used herein, "EC 50 " refers to the concentration of an antibody or antigen-binding portion thereof that induces a response that is 50% of the maximal response (i.e., halfway between the maximal response and the baseline) in either an in vitro or in vivo assay.

[0080] As used herein, the term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve or partially achieve the desired effect. The term "therapeutically effective dose" is defined as an amount sufficient to cure or at least partially halt the disease and its complications in a patient already suffering from the disease. Amounts effective for this use will depend on the severity of the disorder being treated and the general state of the patient's own immune system.

[0081] 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 a process or method of identifying the binding site or epitope of an antibody or antigen-binding fragment thereof on its target protein antigen. Methods and techniques for epitope mapping are provided herein. Epitopes can be formed both 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 on exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost on treatment with denaturing solvents. An epitope typically includes 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, immunoblot 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 those described herein (e.g., X-ray crystallography and 2-dimensional nuclear magnetic resonance) (see, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, GE Morris, Ed. (1996)).

[0082] Antibodies that bind to epitopes on IL-27 that include all or part of the epitopes recognized by the specific antibodies described herein (e.g., the same regions or overlapping regions, or regions between or spanning the regions).

[0083] Antibodies that bind to the same epitope and / or compete with the antibodies described herein for binding to human IL-27 are also covered by the present disclosure. Antibodies that recognize the same epitope or compete for binding can be identified using routine techniques. Such techniques include, for example, immunoassays that show the ability of one antibody to block 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, such as solid-phase direct radioimmunoassays (RIA) or indirect radioimmunoassays, solid-phase direct enzyme immunoassays (EIA) or indirect enzyme immunoassays, sandwich competitive 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 (Cheung et al., Virology 176:546 (1990); and direct label 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 of these, 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. Usually, the test immunoglobulin is present in excess.Typically, when a competing antibody is present in excess, it will inhibit 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.

[0084] Other techniques include epitope mapping methods such as X-ray analysis of crystals of antigen:antibody complexes, which provide atomic resolution of epitopes, and mass spectrometry combined with hydrogen / deuterium (H / D) exchange, which studies the conformation and dynamics of antigen:antibody interactions. Other methods monitor the binding of antibodies to antigen fragments or mutant variants of the antigen, where loss of binding due to modification of amino acid residues within the antigen sequence is often considered indicative of epitope components. In addition, computational combinatorial methods for epitope mapping can also be used. These methods rely on the ability of the antibody of interest to affinity isolate specific short peptides from combinatorial phage display peptide libraries. The peptides are then considered as leads for the definition of 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.

[0085] As used herein, the term "Fc-mediated effector function" or "Fc effector function" refers to the primary function of an antibody and the biological activity of the antibody other than the intended purpose. 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 B cell activation. Many effector functions are initiated by Fc binding to Fcγ receptors. In some embodiments, the tumor antigen-targeting antibody has an effector function (e.g., ADCC activity). In some embodiments, the tumor antigen-targeting antibody described herein comprises a variant constant region that has increased effector function (e.g., increased ability to mediate ADCC) compared to an unmodified form of the constant region.

[0086] As used herein, the term "Fc receptor" refers to a polypeptide found on the surface of immune effector cells and bound by 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 FcRIII (CD16). All four IgG isotypes (IgG1, IgG2, IgG3, and IgG4) bind and activate Fc receptors FcγRI, FcγRIIA, and FcγRIIIA. FcγRIIB is an inhibitory receptor, and thus antibody binding to this receptor does not activate complement and cellular responses. FcγRI is a high affinity receptor that binds IgG in monomeric form, while FcγRIIA and FcγRIIA are low affinity receptors that only bind IgG in multimeric form and have slightly lower affinity. Binding of antibodies to Fc receptors and / or C1q is governed by specific residues or domains within the Fc region. Binding also depends on residues located within the hinge region and within the 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 promoted by the binding of the Fc region to an Fc receptor (e.g., FcγR).

[0087] A list of specific Fc receptor sequences used in this disclosure is provided below as Table 1B.

[0088] As used herein, the term "glycosylation pattern" is defined as the pattern of carbohydrate units covalently attached to a protein, more specifically to an immunoglobulin protein. The glycosylation pattern of a heterologous antibody can be characterized as substantially similar to the glycosylation pattern naturally present on antibodies produced by the non-human transgenic animal species when one skilled in the art would recognize the glycosylation pattern of the heterologous antibody as more similar to the pattern of glycosylation in the non-human transgenic animal species than to the species from which the transgenic CH gene is derived.

[0089] 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 disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation) (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 cover 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).

[0090] As used herein, the term "xenogenous antibody" is defined in relation to the transgenic non-human organism that produces such an antibody. The term refers to an antibody found in an organism that does not constitute a transgenic non-human animal and generally has an amino acid sequence or encoding nucleic acid sequence that corresponds to that from a species other than the transgenic non-human animal species.

[0091] The terms "inducing an immune response" and "promoting an immune response" are used interchangeably and refer to the stimulation of an immune response (i.e., either passive or adaptive) to a particular antigen. The term "inducing," when used in reference to the induction of CDC or ADCC, refers to the stimulation of a specific direct cell killing mechanism.

[0092] As used herein, the term "immunogenic cell death" (alternatively 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 death of the tumor cells 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 mode of immunogenic cell death.

[0093] 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 encompass any measurable decrease 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., 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% inhibition of binding of IL-27.

[0094] As used herein, the terms "inhibiting angiogenesis," "attenuating angiogenesis," and "reducing angiogenesis" refer to reducing the level of angiogenesis in a tissue to an amount that is 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 the same level observed in the control tissue.

[0095] As used herein, the term "inhibit proliferation" (e.g., referring to a cell) is intended to encompass any measurable decrease in proliferation of the cell, e.g., at least about a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100% inhibition of proliferation of the cell.

[0096] As used herein, a "subject in need of prevention," "subject in need of treatment," or "subject in need thereof" refers to a subject who, as determined by a competent medical practitioner (e.g., a physician, nurse, or medical attendant in the case of a human; a veterinarian in the case of a non-human mammal), would reasonably benefit from a given treatment (such as treatment with a composition comprising an anti-IL-27 antibody).

[0097] The term "in vivo" refers to a process that takes place in an organism.

[0098] 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 antigens other than IL-27). However, an isolated antibody that specifically binds to an epitope may have 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. In addition, 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 are combined in a well-defined composition.

[0099] As used herein, the term "isolated nucleic acid molecule" refers to an antibody or antibody portion that binds to IL-27 (e.g., V H , V L, CDR3) of the monoclonal antibody of the anti-IL-27 antibody described herein, and is intended to refer to a nucleic acid molecule in which the nucleotide sequence encoding the antibody or antibody portion is absent other nucleotide sequences encoding antibodies or antibody portions that bind 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 shown in Table 1A may be selected from the sequences shown in Table 1B, which may be a nucleic acid encoding the heavy chain (V H ) and light chain (V L ) corresponds to a nucleotide sequence containing the variable region.

[0100] 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 the disclosure are of the IgG1 isotype. In some embodiments, the human monoclonal antibodies of the disclosure are of the IgG2 isotype. In some embodiments, the human monoclonal antibodies of the disclosure are of the IgG3 isotype. In some embodiments, the human monoclonal antibodies of the disclosure are of the IgG4 isotype. As will be apparent to one of skill in the art, identification of 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 alignments with conserved sequences.

[0101] 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 another.

[0102] 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. D The value of K is the numerical expression of the ratio of the antibody off-rate constant (kd) to the antibody on-rate constant (ka). DThe value of K 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 its antigen. Affinity is the strength of binding of a single molecule to its ligand and is typically measured as the equilibrium dissociation constant (K D ) which is used to assess and rank the strength of orders of magnitude of bimolecular interactions.

[0103] As used herein, "kd" or "k d (Alternatively "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 fraction of the complex that decays or dissociates per second, with units of sec -1 is expressed in

[0104] As used herein, "ka" or "k a (Alternatively "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 expression of the number of antibody / antigen complexes formed per second in a 1 molar (1M) solution of antibody and antigen, and has units of M -1 seconds -1 It is expressed as:

[0105] As used herein, the term "leukocyte" refers to the type of white blood cell involved in defending the body against infectious microorganisms and foreign substances. White blood cells are produced in the bone marrow. There are five main types of white blood cells, subdivided into two main groups: polymorphonuclear leukocytes (neutrophils, eosinophils, basophils) and mononuclear leukocytes (monocytes and lymphocytes).

[0106] As used herein, the term "lymphocyte" refers to a type of leukocyte or white blood cell that is involved in the body's immune defenses. There are two main types of lymphocytes: B cells and T cells.

[0107] As used herein, the terms "linked," "fused," or "fusion" are used interchangeably. These terms refer to the joining together of two or more elements or components or domains by any means, including chemical conjugation or recombinant means. Methods of chemical conjugation (e.g., using heterobifunctional crosslinkers) are known in the art.

[0108] As used herein, "local administration" or "local delivery" refers to the delivery of a composition or agent to its intended target tissue or site that does not depend on transport through the vascular system. For example, the composition can be delivered by injection or implantation of the composition or agent, or by injection or implantation of a device containing the composition or agent. Following local administration near the target tissue or site, the composition or agent, or one or more of its components, can diffuse to the intended target tissue or site.

[0109] 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 may be processed primarily for association with MHC class II. In contrast, antigens delivered endogenously to APCs are processed primarily for association with MHC class I.

[0110] As used herein, the term "monoclonal antibody" refers to an antibody that displays a single binding specificity and affinity for a particular epitope. Thus, the term "human monoclonal antibody" refers to an antibody that displays a single binding specificity and has variable and optional constant regions derived from human germline immunoglobulin sequences. In some embodiments, human monoclonal antibodies are produced by hybridomas that include B cells obtained from a transgenic non-human animal (e.g., a transgenic mouse) whose genome includes human heavy and light chain transgenes, fused to an immortalized cell.

[0111] As used herein, the term "monocyte" refers to a type of white blood cell, which can differentiate into macrophages and dendritic cells to effect an immune response.

[0112] 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, and in antibody-dependent cell-mediated cytotoxicity (ADCC).

[0113] As used herein, the term "naturally occurring" when applied to an object refers to the fact that an object can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism (including viruses) that can be isolated from a natural source and has not been intentionally modified by man in the laboratory is naturally occurring.

[0114] As used herein, the term "nonswitched isotype" refers to the isotype class of heavy chain produced when isotype switching has not occurred, and the CH gene encoding the nonswitched isotype is typically the first CH gene immediately downstream from the functionally rearranged VDJ gene. Isotype switching has been classified as classical or nonclassical isotype switching. Classical isotype switching occurs by recombination events involving at least one switch sequence region in the transgene. Nonclassical isotype switching occurs, for example, in human σ μ and Human Σ μ (δ-associated deletion). Alternative non-classical switching mechanisms (such as inter-transgene and / or inter-chromosomal recombination, among others) may occur to effect isotype switching.

[0115] As used herein, the term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof, either in single-stranded or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogs of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence implicitly encompasses its conservatively modified variants (e.g., degenerate codon substitutions) and complementary sequences in addition to the sequence explicitly indicated. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) 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 at the second base can also be conservative. The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene.

[0116] A polynucleotide as used herein may be composed of any polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA, or modified RNA or DNA. For example, a polynucleotide may 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, and RNA that is a mixture of single-stranded and double-stranded regions, hybrid molecules that include DNA and RNA that may be single-stranded regions or more typically double-stranded regions or mixed single-stranded and double-stranded regions. In addition, a polynucleotide may be composed of RNA or DNA, or triple-stranded regions that include both RNA and DNA. A polynucleotide may also contain one or more modified bases or DNA or RNA backbones that are modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and rare bases (such as inosine). Various modifications can be made to DNA and RNA. Thus, "polynucleotide" encompasses chemically, enzymatically, or metabolically modified forms.

[0117] 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 the sequence. With respect to transcriptional regulatory sequences, operably linked means that the DNA sequences being linked are contiguous, and, where necessary to join two protein coding regions, contiguous in reading frame. With respect to switch sequences, operably linked sequences indicate that switch recombination can be achieved.

[0118] As used herein, "parenteral administration," "administered parenterally," and other grammatically equivalent phrases refer to modes of administration other than enteral and topical administration, usually by injection, including, but 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.

[0119] As used herein, the term "patient" includes human and other mammalian subjects receiving either prophylactic or therapeutic treatment.

[0120] As used herein, the term "PD-1 antagonist" refers to any chemical compound or biological molecule that inhibits the PD-1 signaling pathway or otherwise inhibits PD-1 function in a cell (e.g., an immune cell). In some embodiments, the PD-1 antagonist blocks PD-L1 binding to PD-1 and / or PD-L2 binding 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.

[0121] 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 nucleotides or amino acid residues that are the same when compared and aligned for maximum correspondence, as measured 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 alternatively over the entire length of the two sequences being compared. For 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, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence based on the designated program parameters.

[0122] Optimal alignment of sequences for comparison can be accomplished, 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 computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and 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).

[0123] One example of a suitable algorithm 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 via the website of the National Center for Biotechnology Information.

[0124] Generally as used herein, "pharmacologically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with the tissues, organs, and / or body fluids of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0125] As used herein, "pharmaceutically acceptable carrier" refers to and includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. The composition may include pharmaceutically acceptable salts (e.g., acid addition salts or base addition salts) (see, e.g., Berge et al. (1977) J Pharm Sci 66:1-19).

[0126] As used herein, the terms "polypeptide," "peptide," and "protein" are used interchangeably to refer to a polymer of amino acid residues. These terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.

[0127] As used herein, the term "preventing," when used in reference to a disease state, refers to administration of a composition that reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject compared to a subject who does not receive the composition.

[0128] As used herein, the terms "purified" or "isolated," as applied to any of the proteins (antibodies or fragments) described herein, refer to a polypeptide that has been separated or purified from components (e.g., proteins, or other naturally occurring biological or organic molecules) that naturally associate with the polypeptide (e.g., other proteins, lipids, and nucleic acids in a prokaryote expressing 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.

[0129] As used herein, the term "programmed cell death protein 1" or "PD-1" refers to the programmed cell death protein 1 polypeptide, an immunoinhibitory receptor belonging to the CD28 family, and encoded by the PDCD1 gene in humans. Alternative names or synonyms for PD-1 include PDCD1, PD1, CD279, and SLEB2. PD-1 is preferentially 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" encompasses human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, and analogs that share at least one common epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank accession number AAC51773.

[0130] As used herein, the term "programmed death ligand-1" or "PD-L1" is one of two cell surface glycoprotein ligands for PD-1 (the other is PD-L2) that downregulates T cell activation and cytokine secretion upon binding to PD-1. Alternative names and synonyms for PD-L1 include PDCD1L1, PDL1, B7H1, B7-4, CD274, and B7-H. The term "PD-L1" as used herein encompasses human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, and analogs that share at least one epitope in common with hPD-L1. The complete hPD-L1 sequence can be found under GenBank Accession No. Q9NZQ7.

[0131] PD-1 is known as an immune inhibitory protein that negatively regulates TCR signals (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 between PD-1 and PD-L1 can act as an immune checkpoint, which 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). Immunosuppression can be restored by inhibition of the local interaction of PD-1 with PD-L1 or PD-L2, and the effect is additive when the interaction of PD-L2 with PD-1 is similarly blocked (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).

[0132] For several cancers, tumor survival and proliferation are sustained by tumor-mediated immune checkpoint regulation. This regulation can lead to disruption of anti-cancer immune system function. For example, recent studies have shown that expression of immune checkpoint receptor ligands (such as PD-L1 or PD-L2) by tumor cells can downregulate immune system activity in the tumor microenvironment, particularly by suppressing T cells, and promote cancer immune evasion. PD-L1 is abundantly expressed by various human cancers (Dong et al., (2002) Nat Med 8:787-789). The receptor for PD-L1 (PD-1) is expressed on lymphocytes (e.g., activated T cells) and is usually 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 ligand on tumor cells, the resulting suppression of T cells contributes to a worsening of the immune response against tumors (e.g., reducing tumor-infiltrating lymphocytes or establishing immune evasion by cancer cells).

[0133] For example, in large sample sets of ovarian, renal, colorectal, pancreatic, liver, and melanoma cancers, PD-L1 expression has been shown to correlate with poor prognosis and reduced 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 104:3360-3365). 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. 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 shown 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 via modulation of immune cell-tumor cell interactions.

[0134] As used herein, the term "rearranged" refers to a V segment that is a complete V H Domain or V L Rearranged immunoglobulin gene loci refer to the configuration of a heavy or light chain immunoglobulin locus in which the heavy or light chain immunoglobulin gene loci are positioned immediately adjacent to a DJ segment or J segment, respectively, in a conformation that essentially encodes a rearranged heptamer / nonamer homologous element. Rearranged immunoglobulin gene loci can be identified by comparison to germline DNA, and rearranged loci will have at least one recombined heptamer / nonamer homologous element.

[0135] 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 term is intended to refer not only to the particular subject cell, but also to the progeny of such a cell. Because certain modifications may occur in successive generations due to either mutation or environmental influences, such progeny will not actually be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0136] As used herein, the term "recombinant human antibody" encompasses antibodies prepared, expressed, produced, or isolated by recombinant means, such as (a) antibodies isolated from animals transgenic or transchromosomic for human immunoglobulin genes (e.g., mice) or hybridomas prepared therefrom, (b) antibodies isolated from host cells transformed to express the antibody (e.g., from transfectomas), (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, produced, or isolated by any other means involving splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies are encoded by germline genes that include variable and constant regions that utilize specific human germline immunoglobulin sequences, but include 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), the variable regions contain antigen-binding domains, which are encoded by different genes that rearrange to form antibodies specific to foreign antigens. In addition to rearrangement, the variable regions can be further modified by multiple single amino acid changes (termed somatic mutations or hypermutations) to increase the affinity of the antibody to the foreign antigen. The constant regions will further change in response to the antigen (i.e., isotype switching). Thus, the nucleic acid molecules that are rearranged and somatically mutated in response to antigens, encoding light and heavy immunoglobulin chain polypeptides, may not have sequence identity to the original nucleic acid molecules, but instead will be substantially identical or similar (i.e., have at least 80% identity).

[0137] 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 specific epitope on IL-27, and is used to establish a relationship between the reference antibody itself and one or more distinct antibodies, the relationship being the binding of the reference antibody and one or more distinct antibodies to the same epitope on IL-27. As used herein, the term also connotes an anti-IL-27 antibody that is useful as a competitor in tests or assays such as those described herein (e.g., competitive binding assays), which are useful for discovering, identifying, or developing one or more distinct antibodies that bind to the same epitope.

[0138] As used herein, the terms "specific binding," "selective binding," "selectively binds," and "specifically binds" refer to the binding of an antibody to an epitope on a given antigen. Typically, an antibody binds to an epitope of approximately 10 as determined 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 (approximately 10 -7 , 10 -8 M, 10 -9 M or 10 -10 M or lower in some cases) D ) and binds to a given antigen with an affinity at least 2-fold higher than the affinity for binding to a non-specific antigen other than the given 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 approximately 100 nM (10 -7 M), optionally around 50 nM (5×10 -8 M), optionally approximately 15 nM (1.5×10 -8 M), optionally around 10 nM (10-8 M), optionally less than about 5 nM (5 × 10 -9 M), optionally around 1 nM (10 -9 M), optionally around 0.1 nM (10 -10 M), optionally around 0.01 nM (10 -11 M) or in some cases a lower equilibrium dissociation constant (K D ) and binding to a given antigen occurs with an affinity that is at least two-fold higher than the affinity of the antibody for binding to non-specific antigens other than the given antigen or closely related antigens (e.g., BSA, casein). The phrases "antibody that recognizes an antigen" and "antibody specific for an antigen" are used interchangeably herein with the term "antibody that specifically binds to an antigen."

[0139] As used herein, the term "STAT1 phosphorylation" refers to phosphorylation of the signal transducer and activator of transcription 1 (STAT1) polypeptide (a transcription factor encoded by the STAT1 gene in humans). STAT molecules are phosphorylated by receptor-associated kinases, which cause activation and dimerization by forming homodimers or heterodimers, which translocate to the nucleus and act as transcription factors. STAT1 can be activated (i.e., phosphorylated) in response to signaling through multiple ligands, including IL-27. IL-27 signaling through IL-27R results in phosphorylation of STAT1 (pSTAT1). STAT1 plays an important role in gene expression 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 cytometric analysis of cells labeled with an antibody that specifically recognizes phosphorylated STAT1 (see, e.g., Tochizawa et al., (2006) J Immunol Methods 313(1-2):29-37).

[0140] As used herein, the term "STAT3 phosphorylation" refers to the phosphorylation of signal transducer and activator of transcription 3 (STAT3) polypeptide (a transcription factor encoded by the STAT3 gene in humans). STAT3 mediates the expression of various genes in response to cell stimuli and thus 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 signaling 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).

[0141] As used herein, the term "switch sequence" refers to the DNA sequence responsible for switch recombination. A "switch donor" sequence (typically a μ switch site) will be 5' (i.e., upstream) of the construct region that is deleted during switch recombination. A "switch acceptor" region will be between the construct region to be deleted and the constant region (e.g., gamma, epsilon, etc.) that is replaced. Because there are no specific sites where recombination always occurs, the final gene sequence will typically not be predictable from the construct.

[0142] As used herein, the term "subject" includes any human or non-human animal. For example, the disclosed methods and compositions can be used to treat subjects with immune disorders. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals (non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.).

[0143] With respect to nucleic acids, the term "substantial homology" indicates that two nucleic acids or designated sequences, when optimally aligned and compared, are identical in at least about 80% of the nucleotides, usually at least about 90%-95% and more preferably at least about 98%-99.5% of the nucleotides, with appropriate nucleotide insertions or deletions. Alternatively, substantial homology exists when the segments will hybridize under selective hybridization conditions, to the complement of the strand.

[0144] The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology = number of identical positions / total number of positions x 100), taking into account the number of gaps and the length of each gap that need to be introduced for optimal alignment of the two sequences. As described below in non-limiting examples, the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.

[0145] The percent identity between two nucleotide sequences may be determined using the GAP program in the GCG software package (available at http: / / www.gcg.com) using the NWSgapdna.CMP matrix and gap weights of 40, 50, 60, 70, or 80, and length weights of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide or amino acid sequences may also be determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989)) incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Additionally, the 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 in the GCG software package (available at http: / / www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4, and a length weight of 1, 2, 3, 4, 5, or 6.

[0146] The nucleic acid and protein sequences of the present disclosure can further 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 NBLAST program, score=100, wordlength=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, wordlength=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.

[0147] Nucleic acids may be present in whole cells, in cell lysates, or in partially purified or substantially pure forms. Nucleic acids are "isolated" or "substantially pure" when they are purified from other cellular components or other contaminants (e.g., other cellular nucleic acids or proteins) by standard techniques, including alkaline / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and others known in the art. See F. Ausubel, et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

[0148] The nucleic acid compositions of the present disclosure are often native sequences (except for modified restriction sites and the like), but may be mutated from either cDNA, genome, or mixtures thereof, according to standard techniques for providing gene sequences. For coding sequences, these mutations may affect the amino acid sequence as desired. In particular, DNA sequences that are substantially homologous to or derived from native V, D, J, constant, switch, and other such sequences described herein are contemplated (wherein "derived" indicates that one sequence is identical to or modified from another sequence).

[0149] As used herein, the term "STING" (alternatively TMEM173) refers to stimulator of interferon genes (a protein that functions both as a direct cytoplasmic DNA sensor and an adaptor protein). In humans, STING is encoded by the TMEM173 gene. STING plays an important role in innate immunity. When cells are infected by intracellular pathogens (such as viruses, mycobacteria, and intracellular parasites), STING induces the production of type I interferon. STING-mediated type I interferon protects infected cells and nearby cells from local infection by binding to the same cell that secretes it and nearby cells. An exemplary amino acid sequence for STING is provided by the NCBI Genbank database under accession number NP_001288667.

[0150] 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 T helper cells (also known as T H Cells or CD4 + T cells) and subtypes (T H 1 cell, T H 2 cells, T H 3 cells, T H 17 cells, T H 9 cells, and T FHcells), cytotoxic T cells (also known as T C cells, CD8 + T cells, cytotoxic T lymphocytes, T killer cells, killer T cells), memory T cells and subtypes (central memory T cells (T CM cells) and effector memory T cells (T EM Cells and T EMRA cells), and resident memory T cells (T RM regulatory T cells (also known as T reg cells or suppressor T cells) and subtypes (CD4 + FOXP3 + T reg cells, CD4 + FOXP3 - T reg cells, Tr1 cells, Th3 cells, and T reg 17 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 T cells not mentioned or discussed above may be a target cell type for the methods of use of the present disclosure.

[0151] As used herein, the term "T cell-mediated response" refers to a response that is mediated by effector T cells (e.g., CD8 + cells) and helper T cells (e.g., CD4 + "T-cell mediated responses" refers to any response mediated by T cells, including but not limited to T cells. T cell mediated responses include, for example, T cell cytotoxicity and proliferation.

[0152] 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).

[0153] As used herein, the term "TAM receptor" refers to TAM receptor protein tyrosine kinases (TYRO3, AXL, and MER). TAM receptors are involved in regulating immune system homeostasis. In the context of cancer, TAM receptors have a dual regulatory role: controlling the initiation and progression of tumor development, and simultaneously controlling 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.

[0154] As used herein, the term "TIGIT" or "T cell immunoreceptor with 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 indicated. TIGIT is also known in the art as DKFZp667A205, FLJ39873, V-set immunoglobulin domain-containing protein 9, V-set transmembrane domain-containing protein 3, VSIG9, VSTM3, and WUCAM. The term also encompasses naturally occurring variants (e.g., splice variants or allelic variants) of TIGIT. An exemplary amino acid sequence of human TIGIT can be found under UniProt accession number Q495A1.

[0155] The terms "treat," "treating," and "treatment," as used herein, refer to therapeutic or prophylactic treatments as described herein. Methods of "treatment" employ administering a human antibody of the present disclosure to a subject in need of such treatment (e.g., a subject in need of promoting an immune response against a particular antigen, or a subject who may ultimately acquire such a disorder) to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of a disorder or a recurrent disorder, or to prolong the subject's survival beyond that expected in the absence of such treatment.

[0156] As used herein, the term "tumor microenvironment" (alternatively "cancer microenvironment"; shortened to TME) refers to the environment or milieu in which a tumor or neoplasm resides, including the surrounding blood vessels and cells that contain non-cancerous cells, including but not limited to immune cells, fibroblasts, bone marrow-derived inflammatory cells, and lymphocytes. Signaling molecules and extracellular matrix also comprise the TME. The tumor and the surrounding microenvironment are intimately related and constantly interact. Tumors can affect the microenvironment by releasing extracellular signals, promoting tumor angiogenesis, and inducing peripheral immune tolerance, while immune cells in the microenvironment can affect tumor cell growth and development.

[0157] As used herein, the term "unrearranged" or "germline configuration" refers to a configuration in which the V segment has not been rearranged immediately adjacent to a D or J segment.

[0158] As used herein, the term "vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it is 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) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively 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" can be used interchangeably, as the plasmid is the most commonly used form of vector. However, the disclosure is intended to include other forms of expression vectors that serve equivalent functions, such as viral vectors (eg, replication defective retroviruses, adenoviruses, and adeno-associated viruses).

[0159] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Preferred methods and materials are described below, but methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the methods and compositions disclosed herein. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0160] II. Methods of the Disclosure Some aspects of the present disclosure relate to a method of stimulating an immune response in a subject, comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27 or an antigen-binding portion thereof.Some aspects of the present disclosure relate to a method of treating cancer in a subject in need of treatment, comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27.Some aspects of the present disclosure relate to a method of inhibiting or reducing phosphorylation of STAT1 and / or STAT3 in a cell, comprising contacting the cell with an isolated antibody or antigen-binding fragment provided by the present disclosure, where the antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT1 and / or STAT3 in the cell.Some aspects of the present disclosure relate to a method of inhibiting or reducing inhibition of CD161 expression in a cell, comprising contacting the cell with an isolated antibody or antigen-binding fragment provided by the present disclosure, where the antibody or antigen-binding portion thereof inhibits or reduces inhibition of CD161 expression in the cell. Some aspects of the disclosure relate to a method of inhibiting or reducing expression of PD-L1 in a cell, comprising contacting the cell with an isolated antibody or antigen-binding fragment provided by the disclosure, where the antibody or antigen-binding portion thereof inhibits or reduces expression of PD-L1 in the cell.Some aspects of the disclosure relate to a method of altering TIM-3 expression in a cell, comprising contacting the cell with an isolated antibody or antigen-binding fragment provided by the disclosure, where the antibody or antigen-binding portion thereof alters TIM-3 expression.Some aspects of the disclosure relate to a method of inducing or promoting secretion of one or more cytokines from a cell, comprising contacting the cell with an isolated antibody or antigen-binding fragment provided by the disclosure, where the antibody or antigen-binding portion thereof induces or promotes PD-1 mediated secretion of one or more cytokines from the cell.

[0161] 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 that includes one or more amino acids of (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142-164 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 a pSTAT1 inhibition level IC90, i.e., greater than about 0.7 ug / mL, for the duration of the treatment, e.g., 28 days, 56 days, or 84 days.

[0162] In some embodiments, the antibody or antigen-binding portion thereof is administered in an amount of 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, 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 0.3 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 10 mg / kg, at least about 1 mg / kg to at least about 9 mg / kg, at least about 3 mg / kg 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.

[0163] 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, 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, 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.

[0164] In some embodiments, the antibody or antigen-binding portion thereof is administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 6 weeks, about once every 8 weeks, or about once every 12 weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered about once every 4 weeks.

[0165] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 0.3 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 1 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 2 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 3 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 4 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 5 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 6 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 7 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 8 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 9 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 10 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 11 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 12 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 13 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 14 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 15 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 16 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 17 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 18 mg / kg about once weekly.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 19 mg / kg about once weekly. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 20 mg / kg about once weekly.

[0166] 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.

[0167] 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.

[0168] 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.

[0169] Certain aspects of the present disclosure relate to methods of treating cancer in a subject in need thereof. In some embodiments, the cancer is Kaposi's sarcoma, leukemia, acute lymphocytic leukemia, acute myeloid leukemia, myeloblastic / promyelocytic / myelomonocytic / monocytic erythroleukemia, chronic leukemia, chronic myelogenous (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 lymphoma, or combination therapy with other cancers. Chronic globulinemia, heavy chain disease, solid tumors, sarcomas and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovium, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon 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, bronchogenic carcinoma, Renal cell carcinoma (RCC), hepatocellular carcinoma (HCC), hepatocellular carcinoma, 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 carcinoma, esophageal carcinoma, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system (CN) S) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastric cancer, intraepithelial neoplasia, kidney cancer, laryngeal cancer, liver cancer, lung cancer (small cell, large cell), melanoma, neuroblastoma; oral cancer (e.g., lips, tongue, mouth, and pharynx), ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer; respiratory system cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, urinary tract cancer, 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), colorectal cancer, bladder cancer, endometrial cancer, prostate cancer, thyroid cancer, hepatocellular carcinoma, gastric cancer, brain cancer, 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 therapies for cancer. For example, the composition can be administered to the subject simultaneously with, prior to, or after radiation, surgery, targeted or cytotoxic chemotherapy, chemoradiotherapy, hormone therapy, immunotherapy, gene therapy, cell transplantation therapy, precision medicine, genome editing therapy, or other drug therapy.

[0170] In some embodiments, the compositions disclosed herein are administered to a subject, for example, a human subject, using various methods that depend 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.

[0171] Administration can be achieved, for example, by local infusion, injection, or implantation means. 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 for sustained or periodic release of the composition to the subject. See, for example, U.S. Patent Application Publication No. 20080241223; U.S. Patent Nos. 5,501,856; 4,863,457; and 3,710,795; EP488401; and EP430539, the disclosures of each of which are incorporated herein by reference in their entirety. The composition can be delivered to the subject by means of an implantable device, such as an osmotic pump, a biodegradable implant, an electrodiffusion system, an electroosmotic system, a vapor pressure pump, an electrolytic pump, a foam pump, a piezoelectric pump, a degradable system, or an electromechanical system, for example, by a diffusion system, a decomposition system, or a convection system.

[0172] In some embodiments, the anti-IL-27 antibody or antigen-binding fragment thereof is therapeutically delivered to the subject by local administration techniques.

[0173] In certain embodiments, the route of administration is consistent with known methods, for example, oral, intravenous, intraperitoneal, intracerebral (intracemata), intraventricular, intramuscular, subcutaneous, intraocular, intraarterial, intraportal, or intralesional; by sustained release system, or by implantation device.In certain embodiments, the composition can be administered by bolus injection, by continuous infusion, or by implantation device.In certain embodiments, the individual components of the combination therapy can be administered by different routes.

[0174] In certain embodiments, the composition can be administered locally via implantation of a membrane, sponge, or another suitable material into which the desired molecule is absorbed or encapsulated. In certain embodiments in which an implantation device is used, the device is implanted into any suitable tissue or organ, and delivery of the desired molecule can be via diffusion, sustained release bolus, or continuous administration. In certain embodiments, it may be desirable to use pharmaceutical compositions comprising anti-IL-27 antibodies in an ex vivo manner. In such instances, cells, tissues, and / or organs removed from a patient are exposed to pharmaceutical compositions comprising anti-IL-27 antibodies, after which the cells, tissues, and / or organs are subsequently implanted back into the patient.

[0175] In certain embodiments, anti-IL-27 antibodies can be delivered by implanting specific cells that have been genetically engineered to express and secrete the polypeptide using methods such as those described herein. 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, the cells can be encapsulated to avoid infiltration of surrounding tissues to reduce the chance of an immunological response. In certain embodiments, the encapsulating material is typically a biocompatible semi-permeable polymeric enclosure or membrane that allows release of the protein product(s) but prevents destruction of the cells by the patient's immune system or by other harmful factors from the surrounding tissues.

[0176] 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, and the increased expression of the one or more biomarkers is compared 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), and the increased expression of eotaxin-1 (CCL11) is compared to the expression of eotaxin-1 (CCL11) prior to administration.

[0177] 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 compared to the expression of TARC (CCL17) prior to administration.

[0178] 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 compared to the expression of VEGF-A prior to administration.

[0179] 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 compared to the expression of IL-7 prior to administration.

[0180] 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 compared to the expression of IL-8 prior to administration.

[0181] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of MCP-1, which is compared 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, which is compared to the expression of MCP-4 prior to administration.

[0182] A. Anti-IL-27 Antibodies and Antigen-Binding Portions Thereof Certain aspects of the disclosure relate to methods of administering antibodies, and antigen-binding portions thereof, that specifically bind to IL-27p28 and antagonize IL-27, particularly human IL-27.

[0183] In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces STAT1 and / or STAT3 phosphorylation in a cell of a 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 pSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 25%, at least about 50%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% compared to pSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) prior to administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 90% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) before administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 91% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) before administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 92% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) before administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein).In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 93% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) before administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 94% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) before administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 95% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) before administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 96% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) prior to administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 97% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) prior to administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein).In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 98% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) prior to administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) by at least about 99% compared to PSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling) prior to administration of the antibody or antigen-binding portion thereof (e.g., an anti-IL-27 antibody disclosed herein).

[0184] 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 expression of PD-L1 in a cell of the subject. In some embodiments, the antibody or antigen-binding portion thereof induces or promotes 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 expression of TIM-3 in a cell of the subject. In some embodiments, the cell is a tumor cell or an immune cell.

[0185] Thus, in one aspect, the disclosure provides an isolated antibody, or antigen-binding portion thereof, that specifically binds to and antagonizes human IL-27, and that specifically binds to an epitope disclosed herein and has the following characteristics: (i) an equilibrium dissociation constant (K D(ii) block binding of IL-27 to the IL-27 receptor; (iii) inhibit or reduce phosphorylation of STAT1 and / or STAT3 in a cell; (iv) inhibit or reduce IL-27-mediated inhibition of CD161 expression in a cell; (v) inhibit or reduce IL-27-mediated PD-L1 expression in a cell; (vi) induce or promote PD-1-mediated secretion of one or more cytokines from a cell; (vii) alter expression of TIM-3 in a cell; and (viii) exhibit at least one or more combinations of (i)-(vii).

[0186] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to an epitope that includes one or more amino acids of (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142-164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii). In some embodiments, an isolated antibody or antigen-binding portion thereof of the present disclosure that antagonizes human IL-27 specifically binds to an epitope that includes 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).

[0187] Some embodiments of the present disclosure relate to a method of treating cancer in a subject in need of such treatment, comprising administering to the subject an antibody or antigen-binding portion thereof that antagonizes human IL-27, wherein the antibody or antigen-binding portion thereof specifically binds to an epitope comprising one or more of: (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28); (ii) amino acids 142-164 corresponding to SEQ ID NO:2 (IL-27p28); or (iii) both (i) and (ii), wherein the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.003 mg / kg to at least about 10 mg / kg, and 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 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 antigen-binding 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.

[0188] 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.

[0189] 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.

[0190] 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. In some embodiments, the method comprises administering at least about 0.003 mg / kg to at least about 20 mg / kg of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 1 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 3 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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.In some embodiments, the method comprises administering at least about a 6 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 10 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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.

[0191] 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. In some embodiments, the method comprises administering at least about 0.003 mg / kg to at least about 20 mg / kg of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering a dose of at least about 0.003 mg / kg to at least about 20 mg / kg of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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.In some embodiments, the method comprises administering at least about a 1 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 3 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 6 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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.In some embodiments, the method comprises administering at least about a 10 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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.

[0192] 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.

[0193] 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.

[0194] 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 method comprises administering at least about 0.003 mg / kg to at least about 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about 1 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 3 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 6 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 10 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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. In some embodiments, the method comprises administering at least about a 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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.

[0195] 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 an amino acid sequence set forth in SEQ ID NO:135.

[0196] 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.

[0197] 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.

[0198] 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 method comprises administering at least about 0.003 mg / kg to at least about 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 135, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about 1 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 135, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 3 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 135, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 6 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 135, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 10 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 135, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 135, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137.

[0199] 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. In some embodiments, the method comprises administering at least about 0.003 mg / kg to at least about 20 mg / kg of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about 1 mg / kg of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 3 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 6 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 10 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the method comprises administering at least about a 20 mg / kg dose of an antibody or antigen-binding portion thereof that antagonizes human IL-27, 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: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137.

[0200] In some embodiments, the antibody or antigen-binding portion thereof comprises an amino acid sequence shown in Table 1A. [Table 1A-1] [Table 1A-2] [Table 1A-3] [Table 1A-4] [Table 1A-5] [Table 1A-6] [Table 1A-7] [Table 1A-8] [Table 1A-9] [Table 1A-10] [Table 1A-11] [Table 1A-12] [Table 1A-13] [Table 1A-14] [Table 1A-15] [Table 1A-16]

Table 1A-17

Table 1A-18

Table 1A-19

Table 1A-20

Table 1A-21

Table 1A-22

Table 1A-23

Table 1A-24

Table 1A-25

[0201] In some embodiments, the antibody or antigen-binding portion thereof comprises an Fc sequence as set forth in Table IB. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain, the heavy chain comprising an Fc region having amino acids that have 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, the heavy chain comprising 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, the heavy chain comprising 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, the heavy chain comprising an Fc region comprising the amino acid sequence set forth in SEQ ID NO:7. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain, the heavy chain comprising an Fc region comprising the amino acid sequence set forth in SEQ ID NO:8. [Table 1B]

[0202] 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).

[0203] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope that includes 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 that includes 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).

[0204] 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, 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 disclosure specifically binds to an epitope comprising 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 disclosure specifically binds to an epitope comprising at least one, at least two, at least three, at least four, at least five, 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, 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 including Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO:2 (IL-27p28).

[0205] 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).

[0206] 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).

[0207] 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).

[0208] 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, 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).

[0209] 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, 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).

[0210] In some aspects, 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) and antagonizes human IL-27, and has the following characteristics: (i) an equilibrium dissociation constant (K D(ii) block binding of IL-27 to the IL-27 receptor; (iii) inhibit or reduce phosphorylation of STAT1 and / or STAT3 in the cell; (iv) inhibit or reduce inhibition of CD161 expression in the cell; (v) inhibit or reduce expression of PD-L1 in the cell; (vi) induce or promote PD-1 mediated secretion of one or more cytokines from the cell; (vii) alter expression of TIM-3 in the cell; and (viii) exhibit at least one or more combinations of (i)-(vii).

[0211] In some embodiments, the isolated antibody or antigen-binding portion thereof binds to an epitope comprising 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 ) and binds specifically to it.

[0212] 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.

[0213] In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT1 in a cell. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT3 in a cell. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT1 and STAT3 in a cell. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT1 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 phosphorylation of STAT1 in a 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 phosphorylation of STAT1 in a cell by at least about 50% compared to phosphorylation of STAT1 in a 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 phosphorylation of STAT1 in a cell by at least about 60% compared to phosphorylation of STAT1 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 phosphorylation of STAT1 in a cell by at least about 70% compared to phosphorylation of STAT1 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 phosphorylation of STAT1 in a cell by at least about 75% compared to phosphorylation of STAT1 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 phosphorylation of STAT1 in a cell by at least about 80% compared to phosphorylation of STAT1 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 phosphorylation of STAT1 in a cell by at least about 85% compared to phosphorylation of STAT1 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 phosphorylation of STAT1 in a cell by at least about 90% compared to phosphorylation of STAT1 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 phosphorylation of STAT1 in a cell by at least about 95% compared to phosphorylation of STAT1 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 eliminates phosphorylation of STAT1 in a cell.

[0214] In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT3 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 phosphorylation of STAT3 in a 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 phosphorylation of STAT3 in a cell by at least about 50% compared to phosphorylation of STAT3 in a 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 phosphorylation of STAT3 in a cell by at least about 60% compared to phosphorylation of STAT3 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 phosphorylation of STAT3 in a cell by at least about 70% compared to phosphorylation of STAT3 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 phosphorylation of STAT3 in a cell by at least about 75% compared to phosphorylation of STAT3 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 phosphorylation of STAT3 in a cell by at least about 80% compared to phosphorylation of STAT3 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 phosphorylation of STAT3 in a cell by at least about 85% compared to phosphorylation of STAT3 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 phosphorylation of STAT3 in a cell by at least about 90% compared to phosphorylation of STAT3 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 phosphorylation of STAT3 in a cell by at least about 95% compared to phosphorylation of STAT3 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 eliminates phosphorylation of STAT3 in a cell.

[0215] In some embodiments, the cell is an immune cell. In some embodiments, the cell is a cancer cell.

[0216] 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.

[0217] In some embodiments, the isolated antibody or antigen binding portion thereof inhibits or reduces expression of PD-L1 in a cell. In some embodiments, expression of PD-L1 is inhibited or reduced. In some embodiments, expression of TIM-3 is altered. In some embodiments, both expression of PD-L1 and expression of TIM-3 are altered. In some embodiments, the cell is an immune cell. In some embodiments, the antibody is a monoclonal antibody.

[0218] In some embodiments, the isolated antibody or antigen binding portion thereof induces or promotes 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.

[0219] In some embodiments, the isolated antibody or antigen-binding portion thereof is selected from the group consisting of an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody, an IgM antibody, an IgA1 antibody, an IgA2 antibody, an IgD antibody, and an IgE antibody. 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 mutated IgG4 heavy chain constant region comprises any one of the substitutions S228P, L235E, L235A, or a combination thereof according to EU numbering.

[0220] 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, of any one of the preceding embodiments.

[0221] In some aspects, the 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 an antibody or antigen-binding portion thereof according to any one of the preceding aspects.

[0222] In some aspects, the disclosure provides an isolated antibody or antigen-binding portion thereof, wherein a mutation in 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 both binding to the antibody or antigen-binding portion thereof and to the antibody or antigen-binding portion thereof according to any one of the preceding aspects.

[0223] 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 light chain CDR1 consists of N-XXXXXXLFSSNXKXYXX-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 light chain CDR3 consists of N-XXXASAXXX-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 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.

[0224] 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 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 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 heavy chain CDR2 consists of N-XXSSSXSYXYXXXXXXX-C and heavy chain CDR3 consists of N-XXXXGRTSYTATXHNXXXX-C, wherein X is any amino acid.

[0225] 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 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.

[0226] In some aspects, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope that includes 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), (i) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 9, 10, and 11, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 17, 18, and 19, respectively; (ii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 31, 32, and 33, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 39, 40, and 41, respectively; (iii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 53, 54, and 55, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 61, 62, and 63, respectively; (iv) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 75, 76, and 77, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 83, 84, and 85, respectively; (v) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 97, 98, and 99, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 105, 106, and 107, respectively; or (vi) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 119, 120, and 121, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 127, 128, and 129, respectively; The present invention provides an 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:

[0227] In some aspects, 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), (i) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 9, 10, and 11, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 17, 18, and 19, respectively; (ii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 31, 32, and 33, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 39, 40, and 41, respectively; (iii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 53, 54, and 55, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 61, 62, and 63, respectively; (iv) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 75, 76, and 77, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 83, 84, and 85, respectively; (v) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 97, 98, and 99, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 105, 106, and 107, respectively; or (vi) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 119, 120, and 121, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 127, 128, and 129, respectively; The present invention provides an 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:

[0228] In some aspects, 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), (i) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 9, 10, and 11, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 17, 18, and 19, respectively; (ii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 31, 32, and 33, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 39, 40, and 41, respectively; (iii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 53, 54, and 55, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 61, 62, and 63, respectively; (iv) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 75, 76, and 77, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 83, 84, and 85, respectively; (v) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 97, 98, and 99, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 105, 106, and 107, respectively; or (vi) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 119, 120, and 121, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 127, 128, and 129, respectively; The present invention provides an 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:

[0229] In some aspects, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope that includes 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), (i) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 12, 13, and 14, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 20, 21, and 22, respectively; (ii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 34, 35, and 36, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 42, 43, and 44, respectively; (iii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 56, 57, and 58, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 64, 65, and 66, respectively; (iv) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 78, 79, and 80, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 86, 88, and 89, respectively; (v) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 100, 101, and 102, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 108, 109, and 110, respectively; or (vi) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 122, 123, and 124, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 130, 131, and 132, respectively; The present invention provides an 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:

[0230] In some aspects, 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), (i) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 12, 13, and 14, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 20, 21, and 22, respectively; (ii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 34, 35, and 36, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 42, 43, and 44, respectively; (iii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 56, 57, and 58, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 64, 65, and 66, respectively; (iv) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 78, 79, and 80, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 86, 88, and 89, respectively; (v) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 100, 101, and 102, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 108, 109, and 110, respectively; or (vi) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 122, 123, and 124, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 130, 131, and 132, respectively; The present invention provides an 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:

[0231] In some aspects, 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), (i) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 12, 13, and 14, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 20, 21, and 22, respectively; (ii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 34, 35, and 36, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 42, 43, and 44, respectively; (iii) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 56, 57, and 58, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 64, 65, and 66, respectively; (iv) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 78, 79, and 80, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 86, 88, and 89, respectively; (v) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 100, 101, and 102, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 108, 109, and 110, respectively; or (vi) the sequences of heavy chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 122, 123, and 124, respectively, and the sequences of light chain CDR1, CDR2, and CDR3 as set forth in SEQ ID NOs: 130, 131, and 132, respectively; The present invention provides an 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:

[0232] In some embodiments, the disclosure provides an epitope that includes 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). The present invention provides an isolated antibody or antigen-binding portion thereof that specifically binds to a polypeptide of the present invention, 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 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).

[0233] In some aspects, 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), the antigen-binding portion of which 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).

[0234] In some embodiments, the disclosure provides 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). The present invention provides an isolated antibody or antigen-binding portion thereof that specifically binds to, 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 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).

[0235] In some embodiments, the disclosure provides an antibody that specifically binds to an epitope that includes 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 / or wherein the 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 the heavy chain CDR2 does not comprise N-[G / S]ISSS[S / G][S / A]YI[L / Y]YADSVKG-C (SEQ ID NO: 149).

[0236] In some aspects, 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 , 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).

[0237] In some embodiments, the disclosure provides antibodies that specifically bind 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). The present invention provides 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).

[0238] In some aspects, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope that includes 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), (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 as set forth in SEQ ID NO: 121; and light chain CDR1, CDR2, and CDR3 sequences as 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 as set forth in SEQ ID NO: 124; and a light chain CDR1, CDR2, and CDR3 sequence as set forth in SEQ ID NOs: 130, 131, and 132, respectively; The antibody or antigen-binding portion thereof is provided, which does not comprise:

[0239] In some aspects, 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), (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 as set forth in SEQ ID NO: 121; and light chain CDR1, CDR2, and CDR3 sequences as 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 as set forth in SEQ ID NO: 124; and a light chain CDR1, CDR2, and CDR3 sequence as set forth in SEQ ID NOs: 130, 131, and 132, respectively; The antibody or antigen-binding portion thereof is provided, which does not comprise:

[0240] In some aspects, 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), (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 as set forth in SEQ ID NO: 121; and light chain CDR1, CDR2, and CDR3 sequences as 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 as set forth in SEQ ID NO: 124; and a light chain CDR1, CDR2, and CDR3 sequence as set forth in SEQ ID NOs: 130, 131, and 132, respectively; The antibody or antigen-binding portion thereof is provided, which does not comprise:

[0241] In some aspects, 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), comprising 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 N-QQXXXXP[X] n=0-1 The antibody or antigen-binding portion thereof is provided, each of which does not comprise a light chain CDR3 sequence consisting of: TC (SEQ ID NO: 156).

[0242] In some aspects, 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), comprising 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 N-QQXXXXP[X] n=0-1The antibody or antigen-binding portion thereof is provided, each of which does not comprise a light chain CDR3 sequence consisting of: TC (SEQ ID NO: 156).

[0243] In some aspects, 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 heavy chain CDR1 consists of N-GFTFXXXX-C (SEQ ID NO: 145), 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 N-QQXXXXP[X] n=0-1 The antibody or antigen-binding portion thereof is provided, each of which does not comprise a light chain CDR3 sequence consisting of: TC (SEQ ID NO: 156).

[0244] In some embodiments, the disclosure provides a method for the treatment of IL-27-related disorders comprising administering to a patient a therapeutically effective amount of an IL-27 inhibitor, comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor or 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.

[0245] In some aspects, 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 antibody or antigen-binding portion thereof is not an amino acid sequence selected from the group consisting of:

[0246] In some embodiments, the disclosure provides an epitope that antagonizes IL-27 and includes 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 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.

[0247] In some aspects, 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 antibody or antigen-binding portion thereof does not comprise an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of:

[0248] In some embodiments, the disclosure provides a method for the treatment of IL-27-related disorders comprising administering to the patient a therapeutically effective amount of a compound 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 Glu of SEQ ID NO:2 (IL-27p28). 164, 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.

[0249] In some embodiments, the disclosure provides a method for the treatment of IL-27-related disorders comprising administering to a patient a therapeutically effective amount of an IL-27 inhibitor, comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor, comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor, comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor, comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor, comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor, 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.

[0250] In some embodiments, the present disclosure provides a method for the treatment of IL-27-related disorders comprising administering to the patient a therapeutically effective amount of an IL-27 inhibitor 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 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, wherein 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.

[0251] In some embodiments, the disclosure provides a method for the preparation of IL-27-antagonizing antibodies that antagonize IL-27 and that include 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 wherein the heavy 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: 29, 51, 73, 95, 117, and 139; 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.

[0252] In some aspects, 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, (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 antibody or antigen-binding portion thereof does not comprise an amino acid sequence selected from the group consisting of:

[0253] In some aspects, 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, (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 antibody or antigen-binding portion thereof does not comprise an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of:

[0254] In some aspects, 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, (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 antibody or antigen-binding portion thereof does not comprise an amino acid sequence selected from the group consisting of:

[0255] In some embodiments, the disclosure provides an epitope that antagonizes IL-27 and includes 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). Provided is an isolated antibody or antigen-binding portion thereof that specifically binds to a tope, comprising a heavy chain and a light chain, wherein the heavy chain and the light chain do not contain an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of: (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.

[0256] B. Pharmaceutical Compositions and Formulations In some embodiments, the antibodies or antigen-binding portions thereof useful in the methods and compositions disclosed herein are present in a pharmaceutical composition. Accordingly, some embodiments of the present disclosure relate to a pharmaceutical composition comprising an anti-IL-27 antibody together with a pharma- ceutical acceptable diluent, carrier, solubilizing agent, emulsifying agent, preservative, and / or adjuvant.

[0257] In certain embodiments, acceptable formulation materials are preferably non-toxic to recipients at the dosages and concentrations employed. In certain embodiments, the formulation material(s) are for subcutaneous and / or intravenous administration. In certain embodiments, pharmaceutical compositions contain formulation materials to modify, maintain, or preserve, for example, the pH, osmolality, viscosity, clarity, color, isotonicity, aroma, sterility, stability, rate of dissolution or release, adsorption, or permeation of the composition.In certain embodiments, suitable formulation materials include amino acids (such as glycine, glutamine, asparagine, arginine, or lysine); antimicrobials; antioxidants (such as ascorbic acid, sodium sulfite, or sodium bisulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrate, phosphate, or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, β-cyclodextrin, or hydroxypropyl-β-cyclodextrin); bulking substances; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose, or dextrin); proteins (such as serum albumin, gelatin, or immunoglobulins); colorants, flavorings, and diluents; emulsifiers; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium and the like); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide); solvents (such as glycerin, propylene glycol, or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as Pluronic®, PEG, sorbitan esters, polysorbates (such as polysorbate 20, polysorbate 80, etc.), triton, tromethamine, lecithin, cholesterol, tyloxapol, etc.); stability enhancers (such as sucrose or sorbitol); isotonicity enhancers (such as alkali metal halides, preferably sodium chloride or potassium chloride, mannitol sorbitol, etc.); delivery vehicles; diluents; excipients and / or pharmaceutical adjuvants (Remington's Pharmaceutical Sciences, 18th Edition, A. R. Gennaro, ed., Mack Publishing Company (1995). In certain embodiments, the formulation comprises PBS; 20 mM NaOAC, pH 5.2, 50 mM NaCl; and / or 10 mM NaOAC, pH 5.2, 9% sucrose.In certain embodiments, optimal pharmaceutical compositions can be determined by one of skill in the art depending on, for example, the intended route of administration, delivery format, and desired dosage. See, e.g., Remington's Pharmaceutical Sciences, supra. In certain embodiments, such compositions affect the physical state, stability, rate of in vivo release, and / or rate of in vivo clearance of the anti-IL-27 antibody.

[0258] In certain embodiments, the primary vehicle or carrier in a pharmaceutical composition is either aqueous or non-aqueous in nature. For example, in certain embodiments, a suitable vehicle or carrier is water for injection, saline solution, or artificial cerebrospinal fluid, optionally supplemented with other materials common in compositions for parenteral administration. In certain embodiments, the saline comprises isotonic phosphate buffered saline. In certain embodiments, neutrally buffered saline or saline mixed with serum albumin are further exemplary vehicles. In certain embodiments, the pharmaceutical composition comprises a Tris buffer of about pH 7.0-8.5, or an acetate buffer of about pH 4.0-5.5. In some embodiments, the pharmaceutical composition further comprises sorbitol or a suitable substitute thereof. In certain embodiments, a composition comprising an anti-IL-27 antibody is prepared for storage by mixing a selected composition having a desired degree of purity with optional compounding agents in the form of a lyophilized cake or an aqueous solution (Remington's Pharmaceutical Sciences, supra). Additionally, in certain embodiments, a composition comprising an anti-IL-27 antibody is formulated as a lyophilizate using appropriate excipients (such as sucrose).

[0259] In certain embodiments, pharmaceutical compositions are selected for parenteral delivery. In certain embodiments, compositions are selected for inhalation or delivery via the digestive tract (such as orally). The preparation of such pharma- ceutically acceptable compositions is within the capabilities of those skilled in the art.

[0260] In certain embodiments, the formulation components are present in concentrations that are acceptable to the site of administration. In certain embodiments, a buffer is used to maintain the composition at or slightly below physiological pH, usually within a pH range of about 5 to about 8.

[0261] In certain embodiments, when parenteral administration is intended, the therapeutic composition is in the form of a pyrogen-free parenterally acceptable aqueous solution containing anti-IL-27 antibody in a pharma-ceutically acceptable vehicle.In certain embodiments, the vehicle for parenteral injection is sterile distilled water, in which anti-IL-27 antibody is formulated as a sterile isotonic solution and appropriately stored.In certain embodiments, the preparation includes the formulation of the desired molecule with an agent that can provide controlled or sustained release of the product, such as injectable microspheres, biodegradable particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads, or liposomes, so that the product can be delivered later via depot injection.In certain embodiments, hyaluronic acid is also used.When hyaluronic acid is present, it can have the effect of enhancing the duration in the circulation.In certain embodiments, an implantable drug delivery device is used to introduce the desired molecule.

[0262] In certain embodiments, the pharmaceutical composition is formulated for inhalation. In certain embodiments, the anti-IL-27 antibody is formulated as a dry powder for inhalation. In certain embodiments, the inhalation solution containing the anti-IL-27 antibody is formulated with a propellant for aerosol delivery. In certain embodiments, the solution is nebulized. Pulmonary administration is further described in PCT Application No. PCT / US94 / 001875, which describes pulmonary delivery of chemically modified proteins.

[0263] In certain embodiments, the pharmaceutical compositions disclosed herein are formulated for oral administration. In some embodiments, the pharmaceutical compositions are administered orally. In certain embodiments, the anti-IL-27 antibodies administered in this manner are formulated with or without carriers conventionally used in the compounding of solid dosage forms, such as tablets and capsules. In certain embodiments, the capsules are designed to release the active ingredients of the formulation at the point in the gastrointestinal tract when bioavailability is maximized and degradation prior to systemic circulation is minimized. In certain embodiments, at least one additional agent is included to facilitate absorption of the anti-IL-27 antibodies. In certain embodiments, diluents, flavorings, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents, and binding agents are also utilized.

[0264] In certain embodiments, pharmaceutical compositions contain an effective amount of anti-IL-27 antibody in a mixture with non-toxic excipients suitable for tablet manufacture.In certain embodiments, the solution is prepared in unit dose form by dissolving tablet in sterile water or another suitable vehicle.In certain embodiments, suitable excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate; or binders such as starch, gelatin, or acacia; or lubricants such as magnesium stearate, stearic acid, or talc.

[0265] Additional pharmaceutical compositions, including formulations containing anti-IL-27 antibodies in sustained or controlled delivery formulations, will be apparent to those skilled in the art. In certain embodiments, techniques for formulating various other sustained or controlled delivery means, such as liposome carriers, biodegradable microparticles or porous beads, and depot injections, are also known to those skilled in the art. For example, see PCT Application No. PCT / US93 / 00829, which describes controlled release of porous polymeric microparticles for delivery of pharmaceutical compositions. In certain embodiments, sustained release preparations can include semipermeable polymer matrices in the form of shaped articles, such as films or microcapsules. Sustained release matrices can include polyesters, hydrogels, polylactides (U.S. Pat. No. 3,773,919 and EP 058,481), copolymers of L-glutamic acid and gamma-ethyl L-glutamate (Sidman et al., Biopolymers, 22:547-556 (1983)), poly(2-hydroxyethyl methacrylate) (Langer et al., J. Biomed. Mater. Res., 15:167-277 (1981) and Langer, Chem. Tech., 12:98-105 (1982)), ethylene vinyl acetate (Langer et al., supra), or poly-D(-)-3-hydroxybutyrate (EP 133,988). In certain embodiments, sustained release compositions can also include liposomes, which can be prepared by any of several methods known in the art. See, e.g., Eppstein et al., Proc. Natl. Acad. Sci. USA, 82:3688-3692 (1985); EP 036,676; EP 088,046, and EP 143,949.

[0266] Pharmaceutical compositions used for in vivo administration are typically sterilized. In certain embodiments, this is accomplished by filtration through a sterile filtration membrane. In certain embodiments, in which the composition is lyophilized, sterilization using this method is carried out either before or after lyophilization and reconstitution. In certain embodiments, compositions for parenteral administration are stored in lyophilized form or in a solution. In certain embodiments, parenteral compositions are generally placed into a container with a sterile access port, such as an intravenous solution bag or vial with a stopper that can be pierced by a hypodermic injection needle.

[0267] In certain embodiments, once the pharmaceutical composition has been formulated, it is stored in a sterile vial as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. In certain embodiments, such formulations are stored either in a ready-to-use form or in a form that is reconstituted (e.g., lyophilized) prior to administration. In certain embodiments, kits are provided for making single-dose administration units.In certain embodiments, the kits include both a first container with dried protein and a second container with aqueous formulation.In certain embodiments, the kits include single-chamber and multi-chamber pre-filled syringes (e.g. liquid syringes and lyophilized syringes).

[0268] C. Combination Therapy In some embodiments, the anti-IL-27 antibodies or antigen-binding portions thereof provided by the present disclosure can be used in combination with one or more additional therapeutic agents or treatments, such as another therapeutic agent or treatment for cancer. For example, 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, where the combination provides a therapeutic benefit to the subject having or at risk of developing cancer.

[0269] In some embodiments, the 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 anti-IL-27 antibody, or antigen-binding portion thereof, is administered first in the period of time, and the one or more additional therapeutic agents are administered second (e.g., sequentially) in the period of time. In some embodiments, the one or more additional therapeutic agents are administered first in the period of time, and the anti-IL-27 antibody is administered second in the period of time.

[0270] The anti-IL-27 antibodies or antigen-binding fragments thereof described herein can supplement or enhance previous or current therapy. For example, upon treatment with the anti-IL-27 antibodies or antigen-binding fragments thereof, administration of one or more additional therapeutic agents can be discontinued or reduced, e.g., administered at a lower level. In some embodiments, administration of previous therapy can be maintained. In some embodiments, previous therapy will be maintained until the level of anti-IL-27 antibodies reaches a sufficient level to provide a therapeutic effect.

[0271] In some embodiments, the disclosure provides a method of treating cancer in a subject, comprising administering to the subject an effective amount of an isolated antibody, or antigen-binding portion thereof, provided by the disclosure that specifically binds to and antagonizes IL-27 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 drug, cytotoxic agent, immune-based therapy, cytokine, surgical treatment, radiation treatment, activator of costimulatory molecules, inhibitor of inhibitory molecules, vaccine, or cellular immunotherapy, biologic agent, or combination thereof.

[0272] In some embodiments, the one or more additional therapeutic agents are PD-1 antagonists, TIM-3 inhibitors, LAG-3 inhibitors, TIGIT inhibitors, CD112R inhibitors, TAM inhibitors, STING agonists, 4-1BB agonists, or combinations thereof. In some embodiments, the one or more additional therapeutic agents are CD39 antagonists, CD73 antagonists, CCR8 antagonists, or combinations thereof. In some embodiments, the anti-CD73 is any anti-CD73 antibody disclosed, for example, in U.S. Publication No. 2019 / 0031766 A1, which is incorporated herein by reference in its entirety. In some embodiments, the anti-CD39 is any anti-CD39 antibody disclosed, for example, in International Publication No. WO2019 / 178269 A2, which is incorporated herein by reference in its entirety.

[0273] 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, dimverelimab, 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 inhibitors are selected from the group consisting of FAZ053, atezolizumab, avelumab, durvalumab, and BMS-936559. In some embodiments, the disclosure provides a method of promoting one or more activities of an anti-PD-1 antibody (e.g., promoting PD-1-mediated cytokine secretion from a cell exposed to the anti-PD-1 antibody; promoting anti-PD-1-mediated TNFα secretion; promoting anti-PD-1-mediated IL-6 secretion) comprising exposing a cell to an antibody, or antigen-binding portion thereof, provided by the disclosure, simultaneously or sequentially with the anti-PD-1 antibody, thereby promoting one or more activities of the anti-PD1 antibody.

[0274] 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 (YERVOY®), tremelimumab, pazopanib (VOTRIENT®), and / or rituximab. (Trademark), sorafenib (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 / or spartalizumab.

[0275] In some embodiments, the one or more additional therapeutic agents is a TIM-3 inhibitor, optionally, the TIM-3 inhibitor is MGB453 or TSR-022.

[0276] 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.

[0277] 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.

[0278] 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 antagonist, an A2BR antagonist, or a dual A2AR / A2BR antagonist), a CCR8 antagonist, a CTLA4 antagonist, a VEG-F inhibitor, or a combination thereof.

[0279] 1. Combination with chemotherapy In some embodiments, the methods disclosed herein comprise administering 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, dihydroxyanthracenedione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as analogs or congeners 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-dichlorodiamineplatinum(II) (DDP), protease inhibitors (e.g., ribavirin ... These include rocarbazine, 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)), and antimitotic agents (e.g., vincristine and vinblastine) and temozolomide.

[0280] 2. Combination with PD-1 / PD-L1 antagonist In some embodiments, the methods disclosed herein comprise administering 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 of PD-1 / PD-L1 and / or downstream pathway(s) mediated by human PD-1 / PD-L1 signaling and / or cell processing or other human PD-1 / PD-L1 mediated function.

[0281] Thus, provided herein are PD-1 antagonists that directly or allosterically block, antagonize, inhibit, inhibit, or reduce PD-1 / PD-L1 biological activity, including downstream pathways and / or cellular processes mediated by PD-1 / PD-L1 signaling, such as receptor binding to PD-1 / PD-L1 and / or eliciting a cellular response. Also provided herein are PD-1 antagonists that reduce the quantity or amount of human PD-1 or PD-L1 produced by a cell or a subject.

[0282] In some embodiments, the 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 the mRNA encoding PD-1 or PD-L1 and prevents translation. In some embodiments, the PD-1 antagonist binds to the mRNA encoding PD-1 or PD-L1 and causes degradation and / or turnover.

[0283] In some embodiments, the PD-1 antagonist inhibits PD-1 signaling or function. In some embodiments, the PD-1 antagonist blocks PD-1 binding to PD-L1, PD-L2, or both PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist blocks PD-1 binding to PD-L1. In some embodiments, the PD-1 antagonist blocks PD-1 binding to PD-L2. In some embodiments, the PD-1 antagonist blocks PD-1 binding to both 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.

[0284] In some embodiments, the PD-1 antagonist inhibits binding of PD-1 to its cognate ligand. In some embodiments, the PD-1 antagonist inhibits binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist does not inhibit binding of PD-1 to its cognate ligand.

[0285] 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 specifically 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.

[0286] Several immune checkpoint antagonists, which block or disrupt the interaction between PD-1 and one or both of its ligands, PD-L1 and PD-L2, are in clinical development or currently available to clinicians for the treatment of cancer.

[0287] 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 disclosure include KEYTRUDA® (pembrolizumab, MK-3475, h409A11; see US8952136, US8354509, US8900587, and EP2170959, all of which are incorporated herein by reference in their entireties; Merck), OPDIVO® (nivolumab, BMS-936558, MDX-1106, ONO-4538; see US7595048, US8728474, US9073994, US9067999, EP1537878, US8008449, US8779105, and EP2161336, all of which are incorporated herein by reference in their entireties; Bristol Myers Squibb), MEDI0680 (AMP-514), BGB-A317 and BGB-108 (BeiGene), 244C8 and 388D4 (see WO2016106159, which is incorporated by reference in its entirety; Enumeral Biomedical), PDR001 (Novartis), and REGN2810 (Regeneron). Thus, in some embodiments, the PD-1 antagonist is pembrolizumab. In some embodiments, the PD-1 antagonist is nivolumab. In some embodiments, the methods disclosed herein comprise administering an antibody or antigen-binding portion thereof that specifically binds to IL-27 and pembrolizumab. In some embodiments, the methods disclosed herein comprise administering an antibody or antigen-binding portion thereof that specifically binds to IL-27 and nivolumab.

[0288] Examples of anti-human PD-L1 antibodies or antigen-binding fragments thereof that may comprise a PD-1 antagonist in any of the compositions, methods, and uses provided by the present disclosure include BAVENCIO® (avelumab, MSB0010718C, see WO2013 / 79174, which is incorporated by reference in its entirety; Merck / Pfizer), IMFINZI® (durvalumab, MEDI4736), TECENTRIQ® (VEGF, VEGF-1, VEGF-2, VEGF-3, VEGF-4, VEGF-5, VEGF-6, VEGF-7, VEGF-8, VEGF-9, VEGF-11, VEGF-12, VEGF-13, VEGF-14, VEGF-15, VEGF-16, VEGF-17, VEGF-18, VEGF-19, VEGF-20, VEGF-21, VEGF-22, VEGF-23, VEGF-24, VEGF-25, VEGF-25, VEGF-25, VEGF-26, VEGF-27, VEGF-28, VEGF-29, VEGF-30, VEGF-31, VEGF-32, VEGF-33, VEGF-34, VEGF-35, VEGF-35, VEGF-35, VEGF-35, VEGF-35, VEGF-36, VEGF-37, VEGF-35, VEGF-36, VEGF-37, VEGF-38, VEGF-39 ...9, VEGF-39, VE trademark) (atezolizumab, MPDL3280A, RG7446; see WO2010 / 077634, which is incorporated by reference in its entirety; Roche), MDX-1105 (BMS-936559, 12A4; see US7943743 and WO2013 / 173223, both of which are incorporated by reference in their entirety; 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.

[0289] 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 that contains 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 immunoadhesin molecules 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 a PD-L2-FC fusion protein, AMP-224 (also known as B7-DCIg), that specifically binds to human PD-1.

[0290] It will be appreciated by one of skill in the art 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.

[0291] In some embodiments, the PD-1 / PD-L1 antagonist is a small molecule, a nucleic acid, a peptide, a peptidomimetic, a protein, a carbohydrate, a carbohydrate derivative, or a glycopolymer. Exemplary small molecule PD-1 inhibitors are described in Zhan et al., (2016) Drug Discov Today 21(6):1027-1036.

[0292] 3. Combination with TIM-3 inhibitors In some embodiments, the methods disclosed herein include administering 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.

[0293] 4. Combination with LAG-3 inhibitors In some embodiments, the methods disclosed herein include administering an antibody or antigen-binding portion thereof that specifically binds IL-27 and a LAG-3 inhibitor. In some embodiments, the LAG-3 inhibitor can be 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).

[0294] 5. Other Concomitant Uses In some embodiments, the methods disclosed herein comprise administering 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 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 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 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 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 used in combination (e.g., co-administered) with a tyrosine kinase inhibitor, an agent that targets the adenosine axis (e.g., a CD39 antagonist, a CD73 antagonist, or an A2AR antagonist, A2BR antagonist, or a dual A2AR / A2BR antagonist), a CCR8 antagonist, a CTLA4 antagonist, a VEG-F inhibitor, or a combination thereof.

[0295] In some embodiments, the methods disclosed herein include administering an antibody or antigen-binding portion thereof that specifically binds to IL-27 and a cell therapy. In some embodiments, the cell therapy includes modified immune cell therapy. In some embodiments, the cell therapy includes chimeric antigen receptor (CAR) modified immune cell therapy, e.g., CAR T therapy. In some embodiments, the cell therapy includes engineered T cell receptor (TCR) immune cell therapy. In some embodiments, the cell therapy includes allogeneic tumor infiltrating lymphocyte (TIL) therapy.

[0296] III. Methods for Producing Anti-IL-27 Antibodies and Antigen-Binding Fragments Thereof The present disclosure also features a method for producing any of the anti-IL-27 antibodies or antigen-binding fragments thereof described herein. In some embodiments, the method for preparing the antibodies described herein can include immunizing a subject (e.g., a non-human mammal) with a suitable immunogen. Suitable immunogens for generating any of the antibodies described herein are described herein. For example, to generate an antibody that binds to IL-27p28, one skilled in the art can immunize a suitable subject (e.g., a non-human mammal such as a rat, mouse, gerbil, hamster, dog, cat, pig, goat, horse, or non-human primate) with IL-27. In some embodiments, a full-length human IL-27p28 monomer polypeptide comprising the amino acid sequence shown in SEQ ID NO:2 is used as an immunogen.

[0297] A suitable subject (e.g., a non-human mammal) can be immunized with an appropriate antigen with subsequent boosts sufficient times to induce the production of antibodies by the mammal. The immunogen can be administered to the subject (e.g., a non-human mammal) with an adjuvant. Adjuvants useful in producing antibodies in a subject include, but are not limited to, protein adjuvants; bacterial adjuvants, such as whole bacteria (BCG, Corynebacterium parvum, or Salmonella minnesota) and bacterial components (cell wall skeleton, trehalose dimycolate, monophosphoryl lipid A, methanol extract residue of Mycobacterium tuberculosis (MER), complete or incomplete Freund's adjuvant); viral adjuvants; chemical adjuvants (e.g., aluminum hydroxide, and iodoacetic acid and cholesteryl hemisuccinate). Other adjuvants that can be used in the method to induce an immune response include, for example, cholera toxin and Parapox protein. See also Bieg et al. (1999) Autoimmunity 31(1):15-24. See also, e.g., Lodmell et al. (2000) Vaccine 18:1059-1066; Johnson et al. (1999) J Med Chem 42:4640-4649; Baldridge et al. (1999) Methods 19:103-107; and Gupta et al. (1995) Vaccine 13(14):1263-1276.

[0298] In some embodiments, the method includes preparing a hybridoma cell line that secretes a monoclonal antibody that binds to the immunogen. For example, a suitable mammal (such as a laboratory mouse) is immunized with an IL-27 polypeptide as described above. The antibody-producing cells of the immunized mammal (e.g., splenic B cells) can be isolated 2-4 days after at least one boost of the immunogen and then cultured briefly before being fused with cells of a suitable myeloma cell line. The cells can be fused in the presence of a fusion-promoting agent (e.g., vaccinia virus or polyethylene glycol). The hybrid cells resulting from the fusion are cloned and cell clones secreting the desired antibody are selected. For example, spleen cells of a Balb / c laboratory mouse immunized with a suitable immunogen can be fused with myeloma cell line PAI cells or myeloma cell line Sp2 / 0-Ag14 cells. After fusion, the cells are expanded in a suitable culture medium supplemented at regular intervals with a selective medium (e.g., HAT medium) to prevent normal myeloma cells from outgrowing the desired hybridoma cells. The resulting hybrid cells are then screened for secretion of the desired antibody (e.g., an antibody that binds to human IL-27), and in some embodiments, one of skill in the art can identify anti-IL-27 antibodies from non-immune biased libraries, as described, for example, in U.S. Pat. No. 6,300,064 (Knappik et al.; Morphosys AG) and Schoonbroodt et al. (2005) Nucleic Acids Res 33(9):e81.

[0299] In some embodiments, the methods described herein may include or be used in conjunction with, for example, phage display techniques, bacterial display, yeast surface display, eukaryotic viral display, mammalian cell display, and cell-free (e.g., ribosome display) antibody screening techniques (see, e.g., Etz et al. (2001) J Bacteriol 183:6924-6935; Cornelis (2000) Curr Opin Biotechnol 11:450-454; Klemm et al. (2000) Microbiology 146:3025-3032; Kieke et al. (1997) Protein Eng 10:1303-1310; Yeung et al. (2002) Biotechnol Prog 18:212-220; Boder et al. (2000) Methods Enzymology 328:430-444;Grabherr et al.(2001)Comb Chem High Throughput Screen 4:185-192;Michael et al.(1995)Gene Ther 2:660-668;Pereboev et al.(2001)J Virol 75:7107-7113;Schaffitzel et al.(1999)J Immunol Methods 231:119-135; and Hanes et al. (2000) Nat Biotechnol 18:1287-1292).

[0300] Methods for identifying antibodies using various phage display methods are known in the art. In phage display methods, functional antibody domains are displayed on the surface of phage particles carrying the polynucleotide sequences encoding them. Such phage can be used to display antigen-binding domains of antibodies (such as Fab, Fv, or disulfide bond stabilized Fv antibody fragments) expressed from repertoire or combinatorial antibody libraries (e.g., human or murine). Phages used in these methods are typically filamentous phages (such as fd and M13). Antigen-binding domains are expressed as recombinantly fused proteins to any of the phage coat proteins pIII, pVIII, or pIX. See, e.g., Shi et al. (2010) JMB 397:385-396. Examples of phage display methods that can be used to generate the immunoglobulins or fragments thereof described herein include those described in Brinkman et al. (1995) J Immunol Methods 182:41-50; Ames et al. (1995) J Immunol Methods 184:177-186; Kettleborough et al. (1994) Eur J Immunol 24:952-958; Persic et al. (1997) Gene 187:9-18; Burton et al. (1994) Advances in Immunology 57:191-280; as well as those disclosed in PCT Publication Nos. WO 90 / 02809, WO 91 / 10737, WO 92 / 01047, WO 92 / 18619, WO 93 / 11236, WO 95 / 15982, and WO 95 / 20401. Suitable methods are also described, for example, in U.S. Patent Nos. 5,698,426; 5,223,409; 5,403,484; 5,580,717; 5,427,908; 5,750,753; 5,821,047; 5,571,698; 5,427,908; 5,516,637; 5,780,225; 5,658,727; 5,733,743 and 5,969,108.

[0301] In some embodiments, a phage display antibody library can be generated using mRNA collected from B cells from an immunized mammal. For example, a splenocyte sample containing B cells can be isolated from a laboratory mouse immunized as described above with an IL-27 polypeptide. The mRNA can be isolated from the cells and converted to cDNA using standard molecular biology techniques. See, e.g., Sambrook et al. (1989) "Molecular Cloning: A Laboratory Manual, 2 nd See, for example, "Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane (1988), supra; Benny KC Lo (2004), supra; and Borrebaek (1995), supra. cDNA coding for immunoglobulin heavy and light chain variable region polypeptides are used to construct phage display libraries. Methods for generating such libraries are described, for example, in Merz et al. (1995) J Neurosci Methods 62(1-2):213-9; Di Niro et al. (2005) Biochem J 388(Pt 3):889-894; and Engberg et al. (1995) Methods Mol Biol 51:355-376.

[0302] In some embodiments, a combination of selection and screening can be used to identify an antibody of interest, for example, from a population of hybridoma-derived antibodies or a phage display antibody library. Suitable methods are known in the art and are described, for example, in Hoogenboom (1997) Trends in Biotechnology 15:62-70; Brinkman et al. (1995), supra; Ames et al. (1995), supra; Kettleborough et al. (1994), supra; Persic et al. (1997), supra; and Burton et al. (1994), supra. For example, multiple phagemid vectors, each encoding a fusion protein of a bacteriophage coat protein (e.g., pIII, pVIII, or pIX of M13 phage) and a region combining a different antigen, are produced using standard molecular biology techniques and then introduced into a population of bacteria (e.g., E. coli). Expression of bacteriophage in bacteria may, in some embodiments, require the use of a helper phage. In some embodiments, a helper phage is not required (see, e.g., Chasteen et al., (2006) Nucleic Acids Res 34(21):e145). Phage produced from the bacteria are recovered and then contacted with a target antigen, for example, bound (immobilized) to a solid support. Phage can also be contacted with antigen in solution, with the complex subsequently bound to a solid support.

[0303] Subpopulations of antibodies screened using the above methods can be characterized for their specificity and binding affinity for a particular antigen (e.g., human IL-27p28) using any immunological or biochemical based method known in the art. For example, specific binding of an antibody to IL-27p28 can be determined using, for example, immunological or biochemical based methods, such as, but not limited to, ELISA assays, SPR assays, immunoprecipitation assays, affinity chromatography, and equilibrium dialysis, as described above. Immunoassays that can be used to analyze the immunospecific binding and cross-reactivity of antibodies include, but are not limited to, competitive and non-competitive assay systems, using techniques such as Western blots, RIAs, ELISAs (enzyme-linked immunosorbent assays), "sandwich" immunoassays, immunoprecipitation assays, immunodiffusion assays, agglutination assays, complement fixation assays, immunoradiometric assays, fluorescent immunoassays, and protein A immunoassays. Such assays are routine and well known in the art.

[0304] In embodiments where the selected CDR amino acid sequence is a short sequence (e.g., less than 10-15 amino acids in length), the nucleic acid encoding the CDR can be chemically synthesized, e.g., as described in Shiraishi et al. (2007) Nucleic Acids Symposium Series 51(1):129-130 and U.S. Patent No. 6,995,259. For a given nucleic acid sequence encoding an acceptor antibody, the region of the nucleic acid sequence encoding the CDR can be replaced by a chemically synthesized nucleic acid using standard molecular biology techniques. The 5' and 3' ends of the chemically synthesized nucleic acid can be synthesized to include cohesive end restriction enzyme sites for use in cloning the nucleic acid into the nucleic acid encoding the variable region of the donor antibody.

[0305] In some embodiments, the anti-IL-27 antibodies described herein comprise an altered heavy chain constant region that has reduced effector function (or no effector function) compared to its corresponding unaltered constant region. The effector function involving the constant region of the anti-IL-27 antibody can be modulated by altering the properties of the constant region or Fc region. The altered effector function includes, for example, modulation in one or more of the following activities: antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), apoptosis, binding to one or more Fc receptors, and proinflammatory response. Modulation refers to an increase, decrease, or elimination of the effector function activity exhibited by the subject antibody containing the altered constant region compared to the activity of the form without the constant region alteration. In certain embodiments, modulation encompasses situations where the activity is eliminated or completely absent.

[0306] In one embodiment, the anti-IL-27 antibody described herein comprises an IgG4 heavy chain constant region. In one embodiment, the IgG4 heavy chain constant region is a wild-type IgG4 heavy chain constant region. In another embodiment, the IgG4 constant region comprises a mutation, for example, S228P, and one or both of L235E or L235A according to EU numbering (Kabat, EA, et al., supra). In one embodiment, the anti-IL-27 antibody described herein comprises an IgG1 constant region. In one embodiment, the IgG1 heavy chain constant region is a wild-type IgG1 heavy chain constant region. In another embodiment, the IgG1 heavy chain constant region comprises a mutation.

[0307] An altered constant region with altered FcR binding affinity and / or ADCC activity and / or altered CDC activity is a polypeptide that has either enhanced or diminished FcR binding activity and / or ADCC activity and / or CDC activity compared to the unaltered form of the constant region. An altered constant region that exhibits increased binding to an FcR binds at least one FcR with a higher affinity than the unaltered polypeptide. An altered constant region that exhibits decreased binding to an FcR binds at least one FcR with a lower affinity than the unaltered form of the constant region. Such variants that exhibit reduced binding to FcR may retain little or no appreciable binding to FcR, for example, may bind 0-50% (e.g., less than 50, 49, 48, 47, 46,...

Claims

1. 1. A composition for treating cancer in a subject in need of treatment, comprising an antibody or antigen-binding portion thereof that antagonizes human IL-27, the antibody or antigen-binding portion thereof comprising: (i) amino acids 37-56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142-164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) an epitope comprising one or more amino acids of both (i) and (ii); 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; and The composition is administered at a dose of about 10 mg / kg to about 20 mg / kg.

2. 2. The composition of claim 1, wherein the antibody or antigen-binding portion thereof is administered at a dose of about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, or about 20 mg / kg.

3. 2. The composition of claim 1, wherein the antibody or antigen-binding portion thereof is administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 6 weeks, about once every 8 weeks, or about once every 12 weeks.

4. The antibody or antigen-binding portion thereof, (a) about 10 mg / kg once about every 4 weeks; (b) about 13 mg / kg once about every 4 weeks; (c) about 16 mg / kg once about every 4 weeks; (d) about 20 mg / kg once about every 4 weeks; (e) about 10 mg / kg once about every three weeks; (f) about 13 mg / kg once about every three weeks; (g) about 16 mg / kg once about every three weeks, or (h) about 20 mg / kg once about every three weeks The composition according to claim 1, characterized in that it is administered in a dose of

5. The antibody or antigen-binding portion thereof, (a) inhibiting or reducing IL-27-dependent STAT1 and / or STAT3 phosphorylation in cells of the subject; (b) inhibiting or reducing the inhibition of CD161 expression in cells of the subject; (c) inhibiting or reducing expression of PD-L1 in cells of the subject; (d) inducing or promoting PD-1 mediated secretion of one or more cytokines from cells of said subject; and / or (e) altering expression of TIM-3 in cells of the subject.

6. The composition of claim 5 , wherein the cell is a tumor cell or an immune cell.

7. 2. The composition of claim 1, 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).

8. The epitope is (a) 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), or (b) 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).

2. The composition of claim 1, consisting of or consisting essentially of:

9. 2. The composition of claim 1, 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, and / or 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.

10. The composition of claim 1 , 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.

11. The antibody or antigen-binding portion thereof, (a) 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; or (b) 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. The composition according to any one of claims 1 to 10, comprising:

12. 11. The composition of any one of claims 1 to 10, 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.

13. The composition described in any one of claims 1 to 10, wherein the cancer is non-small cell lung cancer.

14. The composition described in any one of claims 1 to 10, wherein the cancer is hepatocellular carcinoma.

15. The composition described in any one of claims 1 to 10, wherein the cancer is gastric cancer.

16. The composition described in any one of claims 1 to 10, wherein the cancer is renal cell carcinoma.

17. The composition described in any one of claims 1 to 10, wherein the cancer is esophageal cancer.

18. The composition of any one of claims 1 to 10, wherein the antibody or antigen-binding portion thereof is administered to the subject in combination with an additional therapeutic agent.

19. 20. The composition of claim 18, wherein the additional therapeutic agent is administered before, after, or simultaneously with the antibody or antigen-binding portion thereof.

20. 20. The composition of claim 18, wherein the additional therapeutic agent comprises chemotherapy, targeted anti-cancer therapy, oncolytic drugs, cytotoxic agents, immune-based therapy, cytokines, surgical treatment, radiation treatment, activators of costimulatory molecules, inhibitors of inhibitory molecules, vaccines, cellular immunotherapy, biopharmaceuticals, or combinations thereof.

21. 20. The composition of claim 18, 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.

22. The composition of claim 18, wherein the additional therapeutic agent comprises a PD-1 antagonist.

23. 23. The composition of claim 22, 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.

24. The additional therapeutic agent is (a) a PD-L1 inhibitor selected from the group consisting of FAZ053, atezolizumab, avelumab, durvalumab, and BMS-936559; (b) a TIM-3 inhibitor selected from MGB453 or TSR-022; or (c) a LAG-3 inhibitor selected from the group consisting of LAG525, BMS-986016, and TSR-033.

20. The composition of claim 18 comprising:

25. The additional therapeutic agent is 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 (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.

26. Following administration of the antibody or antigen-binding portion thereof, (a) the subject exhibits increased expression of one or more biomarkers selected from the group consisting of EBI3, IL-27, TNFα, MIP-1α (CCL3), IFNγ, IL-10, IL-6, and any combination thereof, wherein said increased expression of said one or more biomarkers is compared to the expression of said one or more biomarkers prior to said administration; (b) the subject exhibits increased expression of EBI3, wherein the increased expression of EBI3 is compared to the expression of EBI3 prior to the administration. (c) 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 said increased expression of said one or more biomarkers is compared to the expression of said one or more biomarkers prior to said administration. (d) the subject exhibits increased expression of Eotaxin-1 (CCL11), wherein said increased expression of Eotaxin-1 (CCL11) is compared to the expression of Eotaxin-1 (CCL11) prior to said administration; and / or (e) the subject exhibits an increased circulating level of IFNγ compared to the circulating level of IFNγ prior to said administration.

Images