Methods of treating cancer using PD-1 axis binding antagonists and tigit inhibitors

Abstract

To provide a medicament for treating cancer in an individual or for delaying progression of cancer.SOLUTION: Provided is a medicament for treating cancer in an individual or for delaying progression of cancer in an individual, the medicament comprising an effective amount of an anti-TIGIT antagonist antibody in combination, wherein the anti-TIGIT antagonist antibody inhibits and / or blocks interaction of TIGIT with CD226 and inhibits interaction of TIGIT with PVR, and the medicament comprises an effective amount of an anti-PD-1 antagonist antibody, wherein the anti-PD-1 antagonist antibody inhibits binding of PD-1 to PD-L1.SELECTED DRAWING: Figure 7A-7C

Description

[Technical Field]

[0001] (Cross-reference to related applications) This application claims priority to U.S. Provisional Application No. 61 / 846,941, filed July 16, 2013, U.S. Provisional Application No. 61 / 865,582, filed August 13, 2013, U.S. Provisional Application No. 61 / 950,754, filed March 10, 2014, U.S. Provisional Application No. 61 / 985,884, filed April 29, 2014, and U.S. Provisional Application No. 61 / 992,109, filed May 12, 2014, each of which is incorporated herein by reference in its entirety.

[0002] (Submit sequence listing as an ASCII text file) The contents of the following submission in an ASCII text file are hereby incorporated by reference in their entirety: Sequence Listing in Computer Readable Format (CRF) (Filename: 146392025940SEQLISTING.TXT, Date Recorded: July 16, 2014, Size: 25KB). [Background technology]

[0003] The provision of two distinct signals to T cells is a widely accepted model for lymphocyte activation of resting T lymphocytes by antigen-presenting cells (APCs). Lafferty et al., Aust. J. Biol. Med. ScL 53:27-42 (1975). This model further provides for self-nonself discrimination and immune tolerance. Bretscher et al., Science 169:1042-1049 (1970); Bretscher, PA, PNAS USA 96:185-190 (1999); Jenkins et al., J. Exp. Med. 165:302-319 (1987). The primary, or antigen-specific, signal is transmitted via the T cell receptor (TCR) after recognition of a foreign antigenic peptide presented in association with the major histocompatibility complex (MHC). Secondary or costimulatory signals are delivered to T cells by costimulatory molecules expressed on antigen-presenting cells (APCs) and induce T cells to promote clonal expansion, cytokine secretion, and effector function. Lenschow et al., Ann. Rev. Immunol. 14:233 (1996). In the absence of costimulation, T cells can become refractory to antigenic stimulation, resulting in a tolerogenic response to either foreign or endogenous antigens.

[0004] In the two-signal model, T cells receive both positive costimulatory and negative inhibitory signals. Regulation of these signals is crucial for maximizing the host's protective immune response while maintaining immune tolerance and preventing autoimmunity. Negative signals appear to be required for the induction of T cell tolerance, while positive signals promote T cell activation.

[0005] Both costimulatory and inhibitory signals are provided to antigen-exposed T cells, and the interplay between costimulatory and inhibitory signals is important in controlling the magnitude of the immune response. Furthermore, the signals provided to T cells change as the infection or immune challenge is cleared, worsens, or persists, and these changes have powerful effects on responding T cells, reshaping the immune response.

[0006] The mechanism of costimulation is of therapeutic interest because manipulation of costimulatory signals has been shown to provide a means to enhance or terminate cell-based immune responses. Recently, it has been discovered that T cell dysfunction or anergy is accompanied by the induced and sustained expression of the inhibitory receptor, programmed death-1 polypeptide (PD-1). Consequently, therapeutic targeting of PD-1 and other molecules that signal through interactions with PD-1, such as programmed death-ligand 1 (PD-L1) and programmed death-ligand 2 (PD-L2), is an area of ​​intense interest.

[0007] PD-L1 is overexpressed in many cancers and is often associated with poor prognosis (Okazaki T et al., Intern. Immun. 2007 19(7):813) (Thompson RH et al., Cancer Res 2006, 66(7):3381). Interestingly, most tumors infiltrating T lymphocytes, unlike normal tissue T lymphocytes and peripheral blood T lymphocytes, predominantly express PD-1, suggesting that upregulation of PD-1 on tumor-reactive T cells may contribute to poor antitumor immune responses (Blood 2009 114(8):1537). This may be due to the exploitation of PD-L1 signaling mediated by PD-L1-expressing tumor cells interacting with PD-1-expressing T cells, resulting in attenuation of T cell activation and evasion of immune surveillance (Sharpe et al., Nat Rev 2002) (Keir ME et al., 2008 Annu. Rev. Immunol. 26:677). Thus, inhibition of PD-L1 / PD-1 interactions may enhance CD8+ T cell-mediated killing of tumors.

[0008] Inhibition of PD-1 axis signaling through its direct ligands (e.g., PD-L1, PD-L2) has been proposed as a means to enhance T cell immunity (e.g., tumor immunity) for the treatment of cancer. Furthermore, a similar enhancement of T cell immunity has been observed by inhibiting the binding of PD-L1 to its binding partner, B7-1. Furthermore, combining inhibition of PD-1 signaling with other signaling pathways that are deregulated in tumor cells may further enhance therapeutic efficacy. There remains a need for optimal therapeutic approaches for the treatment, stabilization, prevention, and / or delay of the onset of various cancers.

[0009] All articles, publications, and patent applications disclosed herein are hereby incorporated by reference in their entirety. Summary of the Invention

[0010] The present invention describes combination therapies comprising a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

[0011] Provided herein is a method for treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

[0012] Also provided herein is a method for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

[0013] Also provided herein is a method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

[0014] Also provided herein is a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

[0015] In some embodiments, the immune-related disease is associated with a T cell dysfunction disorder. In some embodiments, the T cell dysfunction disorder is characterized by a decreased response to antigenic stimulation. In some embodiments, the T cell dysfunction disorder is characterized by T cell anergy or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the T cells are CD4+ and CD8+ T cells. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

[0016] Also provided herein is a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that decreases or inhibits TIGIT expression and / or activity.

[0017] Also provided herein is a method for treating or delaying the progression of cancer in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0018] Also provided herein is a method for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0019] Also provided herein is a method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0020] Also provided herein is a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0021] In some embodiments, the immune-related disease is associated with a T cell dysfunction disorder. In some embodiments, the T cell dysfunction disorder is characterized by a decreased response to antigenic stimulation. In some embodiments, the T cell dysfunction disorder is characterized by T cell anergy, or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the T cells are CD4+ and CD8+ T cells. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

[0022] Also provided herein is a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0023] In some embodiments, an agent that modulates CD226 expression and / or activity can increase and / or stimulate CD226 expression and / or activity.

[0024] In some embodiments, the agent that modulates CD226 expression and / or activity is selected from an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, and an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR.

[0025] In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or antigen-binding fragment thereof.

[0026] In some embodiments, the antagonist of TIGIT expression and / or activity is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide. In some embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or antigen-binding fragment thereof.

[0027] In some embodiments, antagonists of PVR expression and / or activity are small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0028] In some embodiments, agents that inhibit and / or block the interaction of TIGIT with PVR are small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0029] In some embodiments, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVR are small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0030] The present invention also describes combination treatments comprising an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional immune co-inhibitory receptors.

[0031] Provided herein is a method for increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional immune co-inhibitory receptors.

[0032] In some embodiments, the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, TIM3, BTLA, and VISTA. In some embodiments, the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, and TIM3.

[0033] The present invention also describes combination treatments comprising an agent that decreases or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional immune co-stimulatory receptors.

[0034] Provided herein is a method for increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional immune co-stimulatory receptors.

[0035] In some embodiments, the one or more additional immune costimulatory receptors are selected from the group of CD226, OX-40, CD28, CD27, CD137, HVEM, and GITR. In some embodiments, the one or more additional immune costimulatory receptors are selected from the group of CD226, OX-40, CD27, CD137, HVEM, and GITR. In some embodiments, the one or more additional immune costimulatory receptors are selected from the group consisting of OX-40 and CD27.

[0036] In some embodiments, any of the above methods further comprise administering at least one chemotherapeutic agent.

[0037] In some embodiments, the individual in any of the above methods is suffering from cancer. In some embodiments, the individual in any of the above methods is human.

[0038] In some embodiments, the individual's CD4 and / or CD8 T cells have increased or enhanced priming, activation, proliferation, cytokine release and / or cytolytic activity relative to before administration of the combination.

[0039] In some embodiments, the number of CD4 and / or CD8 T cells is increased relative to before administration of the combination. In some embodiments, the number of activated CD4 and / or CD8 T cells is increased relative to before administration of the combination. In some embodiments, the activated CD4 and / or CD8 T cells are increased relative to before administration of the combination. In some embodiments, the activated CD4 and / or CD8 T cells are increased relative to before administration of the combination. + The CD4 and / or CD8 T cells are characterized by increased cytolytic activity relative to prior to administration of the combination. In some embodiments, the CD4 and / or CD8 T cells exhibit increased release of cytokines selected from the group consisting of IFN-γ, TNF-α, and interleukins.

[0040] In some embodiments, the CD4 and / or CD8 T cells are effector memory T cells. In some embodiments, the CD4 and / or CD8 effector memory T cells are γ-IFN-responsive. + In some embodiments, the CD4 and / or CD8 effector memory T cells are characterized by CD44 and / or CD8 T cells and / or enhanced cytolytic activity. high CD62L low The gene is characterized by exhibiting expression of

[0041] In some embodiments, the cancer in any of the above methods has increased levels of T cell infiltration.

[0042] In some embodiments, the agent that reduces or inhibits TIGIT expression and / or activity is selected from the group consisting of antagonists of TIGIT expression and / or activity, antagonists of PVR expression and / or activity, and agents that inhibit interactions and / or intracellular signaling mediated by the binding of TIGIT to PVR.

[0043] In some embodiments, the antagonist of TIGIT expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0044] In some embodiments, the antagonist of PVR expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0045] In some embodiments, the agent that inhibits intracellular signaling mediated by the binding of TIGIT to PVR is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0046] In some embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or antigen-binding fragment thereof.

[0047] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR comprising an amino acid sequence selected from the amino acid sequences: KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6) or RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

[0048] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof is DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13) or DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14) and a light chain comprising the amino acid sequence set forth in

[0049] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof is EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15) or EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16) The heavy chain comprises an amino acid sequence set forth in

[0050] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof is DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13) or DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14) and the antibody heavy chain comprises a light chain comprising the amino acid sequence set forth in EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15) or EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16) The amino acid sequence includes the amino acid sequence described in

[0051] In some embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody is selected from a humanized antibody, a chimeric antibody, a bispecific antibody, a heteroconjugate antibody, and an immunotoxin.

[0052] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR that is at least 90% identical to an HVR set forth in any of KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6), or RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

[0053] In some embodiments, the anti-TIGIT antibody or fragment thereof is DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13) or DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14), or EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15) or EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16) Each of the light chains and / or heavy chains comprises an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in

[0054] In some embodiments, the PD-1 axis binding antagonist is selected from the group consisting of a PD-1 binding antagonist, a PD-L1 binding antagonist, and a PD-L2 binding antagonist.

[0055] In some embodiments, the PD-1 axis binding antagonist is a PD-1 binding antagonist. In some embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to its ligand binding partner. In some embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to PD-L1. In some embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to PD-L2. In some embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to both PD-L1 and PD-L2. In some embodiments, the PD-1 binding antagonist is an antibody. In some embodiments, the PD-1 binding antagonist is MDX-1106 (nivolumab). In some embodiments, the PD-1 binding antagonist is MK-3475 (lambrolizumab). In some embodiments, the PD-1 binding antagonist is CT-011 (pidilizumab). In some embodiments, the PD-1 binding antagonist is AMP-224.

[0056] In some embodiments, the PD-1 axis binding antagonist is a PD-L1 binding antagonist. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1. In some embodiments, the PD-L1 binding antagonist is an antibody.

[0057] In some embodiments, the PD-L1 binding antagonist is selected from the group consisting of YW243.55.S70, MPDL3280A, MDX-1105, and MEDI4736.

[0058] In some embodiments, the anti-PD-L1 antibody comprises a heavy chain comprising the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO: 17), the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO: 19); and a light chain comprising the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO: 20), the HVR-L2 sequence of SASFLYS (SEQ ID NO: 21), and the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO: 22).

[0059] In some embodiments, the anti-PD-L1 antibody is a heavy chain variable region comprising the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23); and a light chain variable region comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24).

[0060] In some embodiments, the PD-1 axis binding antagonist is a PD-L2 binding antagonist. In some embodiments, the PD-L2 binding antagonist is an antibody. In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0061] In some embodiments, the cancer being treated is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic carcinoma, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and other hematological malignancies.

[0062] In some embodiments, the agent that reduces or inhibits TIGIT expression and / or activity is administered continuously. In some embodiments, the agent that reduces or inhibits TIGIT expression and / or activity is administered intermittently. In some embodiments, the agent that reduces or inhibits TIGIT expression and / or activity is administered before the PD-1 axis binding antagonist. In some embodiments, the agent that reduces or inhibits TIGIT expression and / or activity is administered simultaneously with the PD-1 axis binding antagonist. In some embodiments, the agent that reduces or inhibits TIGIT expression and / or activity is administered after the PD-1 axis binding antagonist.

[0063] Also provided herein is a kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

[0064] Also provided herein is a kit comprising a PD-1 axis binding antagonist, an agent that reduces or inhibits TIGIT expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

[0065] Also provided herein is a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with a PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

[0066] Also provided herein is a kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that decreases or inhibits TIGIT expression and / or activity to enhance immune function in an individual with cancer.

[0067] Also provided herein is a kit comprising a PD-1 axis binding antagonist, an agent that reduces or inhibits TIGIT expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual with cancer.

[0068] Also provided herein is a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with a PD-1 axis binding antagonist to enhance immune function in an individual with cancer.

[0069] Also provided herein is a kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that modulates CD226 expression and / or activity to treat or delay the progression of cancer in an individual.

[0070] Also provided herein is a kit comprising a PD-1 axis binding antagonist, an agent that modulates CD226 expression and / or activity, and a package insert containing instructions for using the agent that modulates CD226 expression and / or activity in combination with the PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

[0071] Also provided herein is a kit comprising an agent that modulates CD226 expression and / or activity and a package insert containing instructions for using the agent that modulates CD226 expression and / or activity in combination with a PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

[0072] Also provided herein is a kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that modulates CD226 expression and / or activity to enhance immune function in an individual with cancer.

[0073] Also provided herein is a kit comprising a PD-1 axis binding antagonist, an agent that modulates CD226 expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that modulates CD226 expression and / or activity to enhance immune function in an individual with cancer.

[0074] Also provided herein is a kit comprising an agent that modulates CD226 expression and / or activity and a package insert containing instructions for using the agent that modulates CD226 expression and / or activity in combination with a PD-1 axis binding antagonist to enhance immune function in an individual with cancer.

[0075] In some embodiments, the PD-1 axis binding antagonist comprising the kit is an anti-PD-L1 antibody. In some embodiments, the PD-1 axis binding antagonist comprising the kit is an anti-PD-1 antibody. In some embodiments, the agent that decreases or inhibits TIGIT expression and / or activity comprising the kit is selected from the group consisting of antagonists of TIGIT expression and / or activity, antagonists of PVR expression and / or activity, and agents that inhibit interactions and / or intracellular signaling mediated by the binding of TIGIT to PVR. In some embodiments, the agent that decreases or inhibits TIGIT expression and / or activity comprising the kit is an anti-TIGIT antibody or an antigen-binding fragment thereof.

[0076] In some embodiments, the kit includes an agent that modulates CD226 expression and / or activity, which can increase or stimulate CD226 expression and / or activity. In some embodiments, the agent that modulates CD226 expression and / or activity in the kit is selected from an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, or an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT and / or the antagonist of TIGIT expression and / or activity in the kit is an anti-TIGIT antibody or an antigen-binding fragment thereof.

[0077] In certain aspects, the disclosure provides a method for treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity. In other aspects, the disclosure provides the use of an effective amount of a PD-1 axis-binding antagonist in the manufacture of a medicament for treating or delaying the progression of cancer in an individual, wherein the PD-1 axis-binding agent is used in combination with an agent that reduces or inhibits TIGIT expression and / or activity. In other aspects, the disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for treating or delaying the progression of cancer in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with a PD-1 axis-binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis binding antagonist for use in treating or delaying the progression of cancer in combination with an agent that reduces or inhibits TIGIT expression and / or activity.In another aspect, the disclosure provides a pharmaceutical composition containing an agent that reduces or inhibits TIGIT expression and / or activity for use in treating or delaying the progression of cancer in combination with a PD-1 axis binding antagonist.

[0078] In another aspect, the disclosure provides a method for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis-binding antagonist in the manufacture of a medicament for reducing or inhibiting cancer recurrence or cancer progression in an individual, wherein the PD-1 axis-binding agent is used in combination with an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for reducing or inhibiting cancer recurrence or cancer progression in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with a PD-1 axis-binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis binding antagonist for use in combination with an agent that reduces or inhibits TIGIT expression and / or activity to reduce or inhibit cancer recurrence or cancer progression.In another aspect, the disclosure provides a pharmaceutical composition containing an agent that reduces or inhibits TIGIT expression and / or activity for use in combination with a PD-1 axis binding antagonist to reduce or inhibit cancer recurrence or cancer progression.

[0079] In another aspect, the present disclosure provides a method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the present disclosure provides the use of an effective amount of a PD-1 axis-binding antagonist in the manufacture of a medicament for treating or delaying the progression of an immune-related disease in an individual, wherein the PD-1 axis-binding agent is used in combination with an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the present disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for treating or delaying the progression of an immune-related disease in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with a PD-1 axis-binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition comprising a PD-1 axis binding antagonist for use in treating or delaying the progression of an immune-related disease in combination with an agent that decreases or inhibits TIGIT expression and / or activity.In another aspect, the disclosure provides a pharmaceutical composition comprising an agent that decreases or inhibits TIGIT expression and / or activity for use in treating or delaying the progression of an immune-related disease in combination with a PD-1 axis binding antagonist.

[0080] In another aspect, the disclosure provides a combination comprising an effective amount of a PD-1 axis binding antagonist and an agent that decreases or inhibits TIGIT expression and / or activity.

[0081] In another aspect, the disclosure provides a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis-binding antagonist in the manufacture of a medicament for reducing or inhibiting the progression of an immune-related disease in an individual, wherein the PD-1 axis-binding agent is used in combination with an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for reducing or inhibiting the progression of an immune-related disease in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with a PD-1 axis-binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition comprising a PD-1 axis binding antagonist for use in combination with an agent that reduces or inhibits TIGIT expression and / or activity to reduce or inhibit the progression of an immune-related disease. In another aspect, the disclosure provides a pharmaceutical composition comprising an agent that reduces or inhibits TIGIT expression and / or activity for use in combination with a PD-1 axis binding antagonist to reduce or inhibit the progression of an immune-related disease.

[0082] In certain embodiments that may be combined with any of the preceding embodiments, the immune-related disease is associated with a T cell dysfunction disease. In certain embodiments that may be combined with any of the preceding embodiments, the immune-related disease is a viral infection. In certain embodiments that may be combined with any of the preceding embodiments, the viral infection is a chronic viral infection. In certain embodiments that may be combined with any of the preceding embodiments, the T cell dysfunction disease is characterized by a decreased response to antigenic stimulation. In certain embodiments that may be combined with any of the preceding embodiments, the T cell dysfunction disease is characterized by T cell anergy or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity. In certain embodiments that may be combined with any of the preceding embodiments, the T cell dysfunction disease is characterized by T cell exhaustion. In certain embodiments that may be combined with any of the preceding embodiments, the T cells are CD4+ and CD8+ T cells. In certain embodiments that may be combined with any of the preceding embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

[0083] In another aspect, the disclosure provides a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis-binding antagonist in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the PD-1 axis-binding agent is used in combination with an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with a PD-1 axis-binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition comprising a PD-1 axis binding antagonist for use in combination with an agent that decreases or inhibits TIGIT expression and / or activity to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a pharmaceutical composition comprising an agent that decreases or inhibits TIGIT expression and / or activity for use in combination with a PD-1 axis binding antagonist to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a combination comprising an effective amount of a PD-1 axis binding antagonist and an agent that decreases or inhibits TIGIT expression and / or activity.

[0084] In another aspect, the disclosure provides a method of treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis binding antagonist in the manufacture of a medicament for treating or delaying the progression of cancer in an individual, wherein the PD-1 axis binding agent is used in combination with an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that modulates CD226 expression and / or activity in the manufacture of a medicament for treating or delaying the progression of cancer in an individual, wherein the agent that modulates CD226 expression and / or activity is used in combination with a PD-1 axis binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis binding antagonist for use in combination with an agent that modulates CD226 expression and / or activity to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides pharmaceutical compositions containing an agent that modulates CD226 expression and / or activity for use in combination with a PD-1 axis-binding antagonist to treat or delay the progression of cancer in an individual.

[0085] In another aspect, the disclosure provides a method for reducing or inhibiting cancer recurrence or progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides use of an effective amount of a PD-1 axis binding antagonist in the manufacture of a medicament for reducing or inhibiting cancer recurrence or progression in an individual, wherein the PD-1 axis binding agent is used in combination with an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides use of an effective amount of an agent that modulates CD226 expression and / or activity in the manufacture of a medicament for reducing or inhibiting cancer recurrence or progression in an individual, wherein the agent that modulates CD226 expression and / or activity is used in combination with a PD-1 axis binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis binding antagonist for use in combination with an agent that modulates CD226 expression and / or activity to reduce or inhibit cancer recurrence or progression. In another aspect, the disclosure provides pharmaceutical compositions containing an agent that modulates CD226 expression and / or activity for use in combination with a PD-1 axis-binding antagonist to reduce or inhibit cancer recurrence or cancer progression.

[0086] In another aspect, the disclosure provides a method of treating or delaying the progression of an immune-related disease in an individual, the method comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis binding antagonist in the manufacture of a medicament for treating or delaying the progression of an immune-related disease in an individual, wherein the PD-1 axis binding agent is used in combination with an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that modulates CD226 expression and / or activity in the manufacture of a medicament for treating or delaying the progression of an immune-related disease in an individual, wherein the agent that modulates CD226 expression and / or activity is used in combination with a PD-1 axis binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis binding antagonist for use in combination with an agent that modulates CD226 expression and / or activity to treat or delay the progression of an immune-related disease in an individual. In another aspect, the disclosure provides pharmaceutical compositions containing an agent that modulates CD226 expression and / or activity for use in combination with a PD-1 axis-binding antagonist to treat or delay the progression of an immune-related disease in an individual.

[0087] In another aspect, the disclosure provides a combination comprising an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0088] In another aspect, the disclosure provides a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist and an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis-binding antagonist in the manufacture of a medicament for reducing or inhibiting the progression of an immune-related disease in an individual, wherein the PD-1 axis-binding agent is used in combination with an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that modulates CD226 expression and / or activity in the manufacture of a medicament for reducing or inhibiting the progression of an immune-related disease in an individual, wherein the agent that modulates CD226 expression and / or activity is used in combination with a PD-1 axis-binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis-binding antagonist for use in combination with an agent that modulates CD226 expression and / or activity to reduce or inhibit the progression of an immune-related disease. In another aspect, the disclosure provides pharmaceutical compositions containing an agent that modulates CD226 expression and / or activity for use in combination with a PD-1 axis-binding antagonist to reduce or inhibit the progression of immune-related diseases.

[0089] In certain embodiments that may be combined with any of the preceding embodiments, the immune-related disease is associated with a T cell dysfunction disease. In certain embodiments that may be combined with any of the preceding embodiments, the immune-related disease is a viral infection. In certain embodiments that may be combined with any of the preceding embodiments, the viral infection is a chronic viral infection. In certain embodiments that may be combined with any of the preceding embodiments, the T cell dysfunction disease is characterized by a decreased response to antigenic stimulation. In certain embodiments that may be combined with any of the preceding embodiments, the T cell dysfunction disease is characterized by T cell anergy or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity. In certain embodiments that may be combined with any of the preceding embodiments, the T cell dysfunction disease is characterized by T cell exhaustion. In certain embodiments that may be combined with any of the preceding embodiments, the T cells are CD4+ and CD8+ T cells. In certain embodiments that may be combined with any of the preceding embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

[0090] In another aspect, the disclosure provides a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of a PD-1 axis binding antagonist in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the PD-1 axis binding agent is used in combination with an agent that modulates CD226 expression and / or activity. In another aspect, the disclosure provides the use of an effective amount of an agent that modulates CD226 expression and / or activity in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein an agent that decreases or inhibits CD226 expression and / or activity is used in combination with a PD-1 axis binding antagonist. In another aspect, the disclosure provides a pharmaceutical composition containing a PD-1 axis binding antagonist for use in combination with an agent that modulates CD226 expression and / or activity to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a pharmaceutical composition containing an agent that modulates CD226 expression and / or activity for use in combination with a PD-1 axis binding antagonist to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a combination comprising an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0091] In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates CD226 expression and / or activity. In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates the interaction of CD226 with PVR. In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates intracellular signaling mediated by binding of CD226 to PVR. In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is selected from the group consisting of an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof. In certain embodiments that can be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that inhibits and / or blocks the interaction of CD226 with TIGIT. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or antigen-binding fragment thereof.In certain embodiments that may be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an inhibitory nucleic acid selected from the group consisting of an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera. In certain embodiments that may be combined with any of the preceding embodiments, the antisense polynucleotide targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the interfering RNA targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the catalytic RNA targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the RNA-DNA chimera targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an antagonist of TIGIT expression and / or activity. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or antigen-binding fragment thereof. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is an inhibitory nucleic acid selected from the group consisting of an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of PVR expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that may be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVR is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0092] In another aspect, the present disclosure provides a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional co-inhibitory receptors. In another aspect, the present disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with an agent that reduces or inhibits one or more additional co-inhibitory receptors. In another aspect, the present disclosure provides the use of an effective amount of an agent that reduces or inhibits one or more additional co-inhibitory receptors in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the agent that reduces or inhibits one or more additional co-inhibitory receptors is used in combination with an agent that reduces or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides a pharmaceutical composition containing an agent that reduces or inhibits TIGIT expression and / or activity for use in combination with an agent that reduces or inhibits one or more additional co-inhibitory receptors to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a pharmaceutical composition containing an agent that reduces or inhibits one or more additional co-inhibitory receptors for use in combination with an agent that reduces or inhibits TIGIT expression and / or activity to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a combination comprising an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional co-inhibitory receptors. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional co-stimulatory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, and CD96. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional costimulatory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, and TIM3.

[0093] In another aspect, the present disclosure provides a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional costimulatory receptors. In another aspect, the present disclosure provides the use of an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the agent that reduces or inhibits TIGIT expression and / or activity is used in combination with an agent that increases or activates one or more additional costimulatory receptors. In another aspect, the present disclosure provides the use of an effective amount of an agent that increases or activates one or more additional costimulatory receptors in the manufacture of a medicament for enhancing or stimulating an immune response or function in an individual, wherein the agent that increases or activates one or more additional costimulatory receptors is used in combination with an agent that decreases or inhibits TIGIT expression and / or activity. In another aspect, the disclosure provides a pharmaceutical composition containing an agent that decreases or inhibits TIGIT expression and / or activity for use in combination with an agent that increases or activates one or more additional costimulatory receptors to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a pharmaceutical composition containing an agent that increases or activates one or more additional costimulatory receptors for use in combination with an agent that decreases or inhibits TIGIT expression and / or activity to enhance or stimulate an immune response or function. In another aspect, the disclosure provides a combination comprising an effective amount of an agent that decreases or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional costimulatory receptors. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional costimulatory receptors are selected from the group consisting of CD226, OX-40, CD28, CD27, CD137, HVEM, GITR, MICA, ICOS, NKG2D, and 2B4.In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional costimulatory receptors are selected from the group consisting of CD226, OX-40, CD27, CD137, HVEM, and GITR. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional costimulatory receptors are selected from the group consisting of OX-40 and CD27.

[0094] In certain embodiments that may be combined with any of the preceding embodiments, the method further comprises administering at least one chemotherapeutic agent. In certain embodiments that may be combined with any of the preceding embodiments, the individual is afflicted with cancer. In certain embodiments that may be combined with any of the preceding embodiments, the individual is human. In certain embodiments that may be combined with any of the preceding embodiments, the individual's CD4 and / or CD8 T cells have increased or enhanced priming, activation, proliferation, cytokine release, and / or cytolytic activity relative to before administration of the combination. In certain embodiments that may be combined with any of the preceding embodiments, the number of CD4 and / or CD8 T cells is increased relative to before administration of the combination. In certain embodiments that may be combined with any of the preceding embodiments, the number of activated CD4 and / or CD8 T cells is increased relative to before administration of the combination. In certain embodiments that may be combined with any of the preceding embodiments, the activated CD4 and / or CD8 T cells are activated by γ-IFN. + In certain embodiments that may be combined with any of the preceding embodiments, the CD4 and / or CD8 T cells are characterized by enhanced cytolytic activity relative to prior to administration of the combination. In certain embodiments that may be combined with any of the preceding embodiments, the CD4 and / or CD8 T cells exhibit increased release of cytokines selected from the group consisting of IFN-γ, TNF-α, and interleukins. In certain embodiments that may be combined with any of the preceding embodiments, the CD4 and / or CD8 T cells are effector memory T cells. In certain embodiments that may be combined with any of the preceding embodiments, the CD4 and / or CD8 effector memory T cells exhibit increased release of cytokines selected from the group consisting of IFN-γ, TNF-α, and interleukins. +In certain embodiments that may be combined with any of the preceding embodiments, the CD4 and / or CD8 effector memory T cells are characterized by CD44 high CD62L lowIn certain embodiments that may be combined with any of the preceding embodiments, the cancer has elevated levels of T cell infiltration. In certain embodiments that may be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is selected from the group consisting of an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of PVR expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVR is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3 is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is an inhibitory nucleic acid selected from the group consisting of an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera. In certain embodiments that can be combined with any of the preceding embodiments, the antisense polynucleotide targets TIGIT.

[0095] In certain embodiments that may be combined with any of the preceding embodiments, the interfering RNA targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the catalytic RNA targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the RNA-DNA chimera targets TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or an antigen-binding fragment thereof. In certain embodiments that can be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR comprising an amino acid sequence selected from the amino acid sequence: (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6); or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12). In certain embodiments that can be combined with any of the preceding embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody light chain is In certain embodiments that may be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises the amino acid sequence set forth in: In certain embodiments that may be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises the antibody light chain comprising: TIFF2025186261000004.tif46170, and the antibody heavy chain comprises: TIFF2025186261000005.tif47170. In certain embodiments that may be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof is an antibody selected from the group consisting of a humanized antibody, a chimeric antibody, a bispecific antibody, a heteroconjugate antibody, and an immunotoxin. In certain embodiments that can be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR that is at least 90% identical to an HVR set forth in any one of: (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6); or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12). In certain embodiments that can be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR that is at least 90% identical to an HVR set forth in any one of: (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6). a light chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in TIFF2025186261000006.tif46170; and / or and a heavy chain comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in TIFF2025186261000007.tif47170. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is selected from the group consisting of a PD-1 binding antagonist, a PD-L1 binding antagonist, and a PD-L2 binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is a PD-1 binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to its ligand binding partner. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to PD-L1. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to PD-L2. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist inhibits binding of PD-1 to both PD-L1 and PD-L2. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist is an antibody. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist is MDX-1106. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist is MK-3475. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist is CT-011. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 binding antagonist is AMP-224. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is a PD-L1 binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1.In certain embodiments that may be combined with any of the preceding embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L1 binding antagonist is selected from the group consisting of YW243.55.S70, MPDL3280A, MDX-1105, and MEDI4736. In certain embodiments that may be combined with any of the preceding embodiments, the anti-PD-L1 antibody comprises a heavy chain comprising the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO: 17), the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO: 19); and a light chain comprising the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO: 20), the HVR-L2 sequence of SASFLYS (SEQ ID NO: 21), and the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO: 22). In certain embodiments that may be combined with any of the preceding embodiments, the anti-PD-L1 antibody is EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23), EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23), a heavy chain variable region comprising the amino acid sequence of GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 40), or EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 41);and a light chain variable region comprising the amino acid sequence of: DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24). In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is a PD-L2 binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L2 binding antagonist is an antibody. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L2 binding antagonist is an immunoadhesin. In certain embodiments that can be combined with any of the preceding embodiments, the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic cancer, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and other hematological malignancies. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered continuously. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered intermittently. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered before the PD-1 axis binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered simultaneously with the PD-1 axis binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered after the PD-1 axis binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is administered before the agent that modulates CD226 expression and / or activity.In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is administered simultaneously with the agent that modulates CD226 expression and / or activity. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is administered after the agent that modulates CD226 expression and / or activity. In certain embodiments that may be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered before the agent that decreases or inhibits one or more additional immune co-inhibitory receptors. In certain embodiments that may be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered simultaneously with the agent that decreases or inhibits one or more additional immune co-inhibitory receptors. In certain embodiments that may be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered after the agent that decreases or inhibits one or more additional immune co-inhibitory receptors. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered before the agent that increases or activates one or more additional immune costimulatory receptors. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered simultaneously with the agent that increases or activates one or more additional immune costimulatory receptors. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is administered after the agent that increases or activates one or more additional immune costimulatory receptors.

[0096] In other aspects, the disclosure provides kits comprising a PD-1 axis binding antagonist and a package insert comprising instructions for using the PD-1 axis binding antagonist in combination with an agent that decreases or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

[0097] In another aspect, the disclosure provides a kit comprising a PD-1 axis binding antagonist, an agent that reduces or inhibits TIGIT expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

[0098] In another aspect, the present disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert comprising instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with a PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

[0099] In another aspect, the disclosure provides a kit comprising a PD-1 axis binding antagonist and a package insert comprising instructions for using the PD-1 axis binding antagonist in combination with an agent that decreases or inhibits TIGIT expression and / or activity to enhance immune function in an individual with cancer.

[0100] In another aspect, the present disclosure provides a kit comprising a PD-1 axis binding antagonist, an agent that reduces or inhibits TIGIT expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual with cancer.

[0101] In another aspect, the present disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with a PD-1 axis binding antagonist to enhance immune function in an individual with cancer.

[0102] In other aspects, the disclosure provides kits comprising a PD-1 axis binding antagonist and a package insert comprising instructions for using the PD-1 axis binding antagonist in combination with an agent that modulates CD226 expression and / or activity to treat or delay the progression of cancer in an individual.

[0103] In other aspects, the disclosure provides kits comprising a PD-1 axis binding antagonist, an agent that modulates CD226 expression and / or activity, and a package insert comprising instructions for using the agent that modulates CD226 expression and / or activity in combination with the PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

[0104] In other aspects, the present disclosure provides kits comprising an agent that modulates CD226 expression and / or activity and a package insert comprising instructions for using the agent that modulates CD226 expression and / or activity in combination with a PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

[0105] In another aspect, the disclosure provides a kit comprising a PD-1 axis binding antagonist and a package insert comprising instructions for using the PD-1 axis binding antagonist in combination with an agent that modulates CD226 expression and / or activity to enhance immune function in an individual with cancer.

[0106] In another aspect, the present disclosure provides a kit comprising a PD-1 axis binding antagonist, an agent that modulates CD226 expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that modulates CD226 expression and / or activity to enhance immune function in an individual with cancer.

[0107] In another aspect, the present disclosure provides a kit comprising an agent that modulates CD226 expression and / or activity and a package insert containing instructions for using the agent that modulates CD226 expression and / or activity in combination with a PD-1 axis binding antagonist to enhance immune function in an individual with cancer.

[0108] In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis-binding antagonist is an anti-PD-L1 antibody. In certain embodiments that may be combined with any of the preceding embodiments, the anti-PD-L1 antibody is selected from the group consisting of YW243.55.S70, MPDL3280A, MDX-1105, and MEDI4736. In certain embodiments that may be combined with any of the preceding embodiments, the anti-PD-L1 antibody comprises a heavy chain comprising the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO: 17), the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO: 19); and a light chain comprising the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO: 20), the HVR-L2 sequence of SASFLYS (SEQ ID NO: 21), and the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO: 22). In certain embodiments that may be combined with any of the preceding embodiments, the anti-PD-L1 antibody comprises: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23), EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 40), or EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 41) a heavy chain variable region comprising the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24) and a light chain variable region comprising the amino acid sequence of: In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is an anti-PD-1 antibody. In certain embodiments that may be combined with any of the preceding embodiments, the anti-PD-1 antibody is MDX-1106, MK-3475, or CT-011. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is AMP-224. In certain embodiments that may be combined with any of the preceding embodiments, the PD-1 axis binding antagonist is a PD-L2 binding antagonist. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L2 binding antagonist is an antibody. In certain embodiments that may be combined with any of the preceding embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0109] In another aspect, the disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert comprising instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that reduces or inhibits one or more additional immune co-inhibitory receptors to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional immune co-inhibitory receptors, and a package insert comprising instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that reduces or inhibits one or more additional immune co-inhibitory receptors to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides a kit comprising an agent that reduces or inhibits one or more additional immune co-inhibitory receptors and a package insert containing instructions for using the agent that reduces or inhibits one or more additional immune co-inhibitory receptors in combination with an agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that reduces or inhibits one or more additional immune co-inhibitory receptors to enhance immune function in an individual afflicted with cancer. In another aspect, the present disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity, an agent that reduces or inhibits one or more additional immune co-inhibitory receptors, and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that reduces or inhibits one or more additional immune co-inhibitory receptors to enhance immune function in an individual with cancer.In another aspect, the disclosure provides kits comprising an agent that reduces or inhibits one or more additional immune co-inhibitory receptors and a package insert comprising instructions for using the agent that reduces or inhibits one or more additional immune co-inhibitory receptors in combination with an agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual afflicted with cancer. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional costimulatory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, and CD96. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional costimulatory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, and TIM3.

[0110] In another aspect, the disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert comprising instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that increases or activates one or more additional immune costimulatory receptors to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional immune costimulatory receptors, and a package insert comprising instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that increases or activates one or more additional immune costimulatory receptors to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides a kit comprising an agent that increases or activates one or more additional immune costimulatory receptors and a package insert containing instructions for using the agent that increases or activates one or more additional immune costimulatory receptors in combination with an agent that decreases or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual. In another aspect, the disclosure provides a kit comprising an agent that decreases or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that decreases or inhibits TIGIT expression and / or activity in combination with an agent that increases or activates one or more additional immune costimulatory receptors to enhance immune function in an individual with cancer. In another aspect, the present disclosure provides a kit comprising an agent that reduces or inhibits TIGIT expression and / or activity, an agent that increases or activates one or more additional immune costimulatory receptors, and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that increases or activates one or more additional immune costimulatory receptors to enhance immune function in an individual suffering from cancer.In another aspect, the disclosure provides a kit comprising an agent that upregulates or activates one or more additional immune costimulatory receptors and a package insert comprising instructions for using the agent that upregulates or activates one or more additional immune costimulatory receptors in combination with an agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual with cancer. In certain embodiments that can be combined with any of the preceding embodiments, the one or more additional immune costimulatory receptors are selected from the group consisting of CD226, OX-40, CD28, CD27, CD137, HVEM, GITR, MICA, ICOS, NKG2D, and 2B4. In certain embodiments that can be combined with any of the preceding embodiments, the one or more additional immune costimulatory receptors are selected from the group consisting of CD226, OX-40, CD27, CD137, HVEM, and GITR. In certain embodiments that may be combined with any of the preceding embodiments, the one or more additional immune costimulatory receptors are selected from the group consisting of OX-40 and CD27.

[0111] In certain embodiments that can be combined with any of the preceding embodiments, the individual is a human. In certain embodiments that can be combined with any of the preceding embodiments, the agent that decreases or inhibits TIGIT expression and / or activity is selected from the group consisting of an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, and an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3. In certain embodiments that can be combined with any of the preceding embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or antigen-binding fragment thereof. In certain embodiments that can be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates CD226 expression and / or activity. In certain embodiments that can be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates the interaction of CD226 with PVR. In certain embodiments that can be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates intracellular signaling mediated by CD226 binding to PVR.In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is selected from the group consisting of an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, and an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3. In certain embodiments that may be combined with any of the preceding embodiments, the agent that modulates CD226 expression and / or activity is an agent that inhibits and / or blocks the interaction of CD226 with TIGIT. In certain embodiments that may be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide. In certain embodiments that may be combined with any of the preceding embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or antigen-binding fragment thereof.In certain embodiments that can be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR comprising an amino acid sequence selected from the amino acid sequence: (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6); or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12). In certain embodiments that can be combined with any of the preceding embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody light chain is In certain embodiments that may be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises the antibody heavy chain comprising: In certain embodiments that may be combined with any of the preceding embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises the antibody light chain comprising: TIFF2025186261000010.tif47170, and the antibody heavy chain comprises: Contains the amino acid sequence described in TIFF2025186261000011.tif46170. [Brief explanation of the drawings]

[0112] [Figure 1A-1B]Figure 1 shows that TIGIT is highly expressed on exhausted CD8+ and CD4+ T cells. Figure 1A shows MACS-enriched C57BL6 / J splenic CD8+ T cells stimulated with plate-bound anti-CD3 and anti-CD28 for 24–48 h in vitro. Flow cytometry histograms show TIGIT expression (red) relative to isotype staining (gray). Quantification of TIGIT MFI is also shown. ***, P<0.001. Data are representative of two independent experiments; n=3. In Figures 1B–1C, C57BL6 / J mice were infected with Armstrong strain LCMV, and splenocytes were analyzed 7 days postinfection. Data are representative of two independent experiments; n=5. Figure 1B shows flow cytometry histograms depicting TIGIT expression by naive (CD44lowCD62Lhigh) and effector memory (CD44highCD62Llow) CD4+ and CD8+ T cells. Quantification of TIGIT MFI is also shown. ***, P<0.001. [Figure 1C-1D] Figure 1C shows flow cytometry histograms depicting TIGIT expression by PD-1 high and PD-1 low effector memory CD8+ T cells. Quantification of TIGIT MFI is also shown. ***, P<0.001. Figure 1D shows C57BL6 / J mice were acutely depleted of CD4+ T cells and infected with clone 13 strain LCMV. Splenocytes were analyzed 42 days postinfection. Flow cytometry histograms depict TIGIT expression by naive (CD44 low CD62 L high), central memory (CD44 high CD62 L high), and effector memory (CD44 high CD62 L low) CD8+ T cells. Quantification of TIGIT MFI is also shown. ***, P<0.001. Data are representative of two independent experiments; n=5. Error bars indicate the standard error of the mean.

[0113] [Figure 2] Figure 2 shows the design of the TIGIT loxP / loxP mouse. Exon 1 of TIGIT has been flanked with loxP sites using standard techniques.

[0114] [Figure 3] Figure 3 shows that TIGIT-deficient CD8+ and CD4+ T cells respond normally to acute viral infection. TIGITfl / flCD4cre (CKO) and TIGITfl / fl littermates (WT) were infected with Armstrong strain LCMV. Splenocytes were analyzed 7 days postinfection. Data are representative of two independent experiments; n = 5. Figure 3A shows a representative FACS plot gated on CD8+ T cells; activated (CD44high) cells are boxed. Activated CD8+ T cells are quantified as a percentage of total CD8+ T cells. Figure 3B shows a representative FACS plot gated on CD8+ T cells after in vitro stimulation; IFNγ-producing cells are boxed. IFNγ-producing cells are quantified as a percentage of total CD8+ T cells. Figure 3C shows a representative FACS plot gated on CD4+ T cells; activated (CD44high) cells are boxed. Quantification of activated CD4+ T cells as a percentage of total CD4+ T cells. Figure 3D shows a representative FACS plot gated on CD4+ T cells after in vitro stimulation, with IFNγ-producing cells boxed. Quantification of IFNγ-producing cells as a percentage of total CD4+ T cells. Error bars indicate standard error of the mean.

[0115] [Figures 4A-4D]Figure 4 shows that TIGIT and PD-1 synergistically regulate exhausted T cell effector function in vivo. In Figures 4A-4E, TIGITfl / flCD4-cre- (WT) and TIGITfl / flCD4-cre+ (CKO) mice were acutely depleted of CD4+ T cells and infected with clone 13 LCMV. Splenocyte and liver viral titers were analyzed 42 days postinfection. Data are representative of two independent experiments, n = 6-9 per group. Figure 4A shows a representative FACS plot gated on CD8+ T cells; activated cells (CD44highCD62low) are boxed. Activated cells are quantified as a percentage of total CD8+ T cells. Figure 4B shows a representative FACS plot gated on CD8+ T cells after in vitro stimulation; IFNγ+ cells are boxed. Quantification of IFNγ-producing cells as a percentage of CD8+ T cells. Figure 4C shows a representative FACS plot gated on CD4+ T cells, with activated cells (CD44highCD62Llow) boxed. Quantification of activated cells as a percentage of total CD4+ T cells. Figure 4D shows a representative FACS plot gated on CD4+ T cells after in vitro stimulation, with IFNγ+ cells boxed. Quantification of IFNγ-producing cells as a percentage of total CD4+ T cells. [Figures 4E-4F]Figure 4E shows quantification of liver LCMV titers. ***, P<0.0001. In Figures 4F-4H, C57BL6 / J mice were acutely depleted of CD4+ T cells and infected with clone 13 LCMV. Mice were treated with isotype-matched control, anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT antibodies starting 28 days post-infection. Splenocyte and liver viral titers were analyzed 42 days post-infection. Data are representative of two independent experiments; n=10. Figure 4F shows a representative FACS plot gated on CD8+ T cells; activated cells (CD44highCD62Llow) are boxed. Activated cells are quantified as a percentage of total CD8+ T cells. ***, P<0.0001. [Figures 4G-4H] Figure 4G shows a representative FACS plot gated on activated CD8+ T cells after in vitro stimulation; IFNγ+ cells are boxed. Quantification of IFNγ-producing cells is as a percentage of activated CD8+ T cells. *, P = 0.0352. **, P = 0.0047. Figure 4H shows quantification of liver LCMV titers. *, P = 0.0106. **, P = 0.0047. Error bars represent standard error of the mean.

[0116] [Figure 5]Figure 5 shows that TIGIT / PD-L1 co-blockade enhances CD4+ T cell effector function during chronic viral infection. C57BL6 / J mice were depleted of CD4+ T cells and infected with clone 13 LCMV. Mice were treated with isotype control, anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT antibodies starting 28 days post-infection. Splenocyte and liver viral titers were analyzed 42 days post-infection. Data are representative of two independent experiments; n = 10. Figure 5A shows a representative FACS plot gated on CD4+ T cells; activated cells (CD44highCD62Llow) are boxed. Quantification of activated CD4+ T cells is as a percentage of total CD4+ T cells. Figure 5B shows a representative FACS plot gated on CD4+ T cells after in vitro stimulation; IFNγ+ cells are boxed. Quantification of IFNγ-producing cells as a percentage of total CD4+ T cells. *, P=0.019. Error bars indicate standard error of the mean.

[0117] [Figures 6A-6B] Figure 6 shows that TIGIT expression is elevated in human breast cancer and correlates with the expression of CD8 and inhibitory co-receptors. Breast cancer gene expression microarray data generated by the Cancer Gene Atlas Network were analyzed. Gene expression data were normalized and expressed as relative ratios (log2). Figure 6A shows TIGIT expression in normal and whole breast tumor samples (left) and breast tumor subtypes (right). ***, P = 6 × 10-12. Box and whisker plots are shown. Figure 6B shows the correlation between TIGIT and CD3ε expression. R2 = 0.61. [Figure 6C-6D] Figure 6C shows the correlation between TIGIT and CD8α (left, R2 = 0.80) or CD4 (right, R2 = 0.42). Figure 6D shows the correlation between TIGIT and PD-1 (left, R2 = 0.87), LAG3 (center, R2 = 0.80), and CTLA4 (right, R2 = 0.76).

[0118] [Figure 7A-7C]Figure 7 shows that TIGIT and PD-1 inhibit antitumor T cell responses. In Figures 7A-7B, BALB / C mice were inoculated with CT26 colorectal cancer cells. Splenocytes and tumor-infiltrating lymphocytes (TILs) were analyzed 14 days after inoculation, when tumors had reached a size of approximately 200 mm. Data are representative of one experiment; n = 6. Figure 7A shows flow cytometry histograms representing TIGIT expression by spleen- and tumor-infiltrating CD8+ T cells. Quantification of TIGIT MFI is also shown. **, P = 0.0023. Figure 7B shows flow cytometry histograms representing TIGIT expression by spleen- and tumor-infiltrating CD4+ T cells. Quantification of TIGIT MFI is also shown. ***, P = 0.0002. In Figures 7C-7E, BALB / C mice were inoculated with CT26 colorectal cancer cells. When tumors reached approximately 200 mm in size, mice were treated with isotype control, anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT antibodies for 3 weeks. Data are representative of two independent experiments; n = 10-20 (Figures 7C-7D) or 7-10 (Figure 7E). Figure 7C shows the median CT26 tumor volume over time. [Figure 7D] FIG. 7D shows the survival time of the mice. [Figure 7E] Figure 7E shows mice in complete remission (CR) treated with anti-TIGIT plus anti-PD-L1 and naive BALB / c mice inoculated with CT26 cells in the left thoracic flank and EMT6 breast cancer cells in the mammary fat pad approximately 60 days after the first inoculation. Median (left) and individual (right) tumor volumes of CT26 (rectangles) and EMT6 (triangles) tumors in CR mice (purple and green) and naive mice (black and orange) are shown. [Figure 7F]Figure 7F shows mice were inoculated with CT26 tumors and treated as in Figure 7C. Tumor-infiltrating and tumor-draining lymph node-resident T cells were analyzed by flow cytometry. Representative FACS plots of CD8+ TILs after in vitro stimulation; IFNγ-producing cells are boxed. Quantification of IFNγ-producing CD8+ TILs as a percentage of total CD8+ TILs. ***, P=0.0003. Data are representative of two independent experiments; n=5. Error bars indicate standard error of the mean.

[0119] [Figure 8] Figure 8 shows that CT26 tumor-infiltrating lymphocyte TIGIT expression correlates with Tim-3 expression. BALB / C mice were inoculated with CT26 colorectal cancer cells. Splenocytes and tumor-infiltrating lymphocytes (TILs) were analyzed approximately 14 days after inoculation, when tumors had reached a size of approximately 200 mm. Data are representative of one experiment; n = 6. Figure 8A shows representative histograms of TIGIT expression by spleen- and tumor-infiltrating CD8+ T cells. Quantification of TIGIT MFI. **, P = 0.0026. Figure 8B shows representative histograms of TIGIT expression by spleen- and tumor-infiltrating CD4+ T cells. Quantification of TIGIT MFI. ***, P < 0.0001. Error bars indicate standard error of the mean.

[0120] [Figure 9]Figure 9 shows that MC38 tumor-infiltrating lymphocyte TIGIT expression correlates with PD-1 and Tim-3 expression. C57BL6 / J mice were inoculated with MC38 colorectal cancer cells. Splenocytes and tumor-infiltrating lymphocytes (TILs) were analyzed approximately 14 days after inoculation, when tumors had reached a size of approximately 200 mm. Data are representative of one experiment; n = 5. Figure 9A shows representative histograms of TIGIT expression by spleen- and tumor-infiltrating CD8+ T cells. Quantification of TIGIT MFI. ***, P < 0.0001. Figure 9B shows representative histograms of TIGIT expression by spleen- and tumor-infiltrating CD4+ T cells. Quantification of TIGIT MFI. *, P = 0.0136. **, P = 0.0029. Error bars indicate standard error of the mean.

[0121] [Figure 10] Figure 10 shows CT26 tumor growth in mice treated with anti-PD-L1 and / or anti-TIGIT. Naive BALB / c mice were inoculated with CT26 tumor cells and treated with anti-PD-L1 and / or anti-TIGIT or isotype-matched control antibodies as described in Figures 4D-4F. Tumor volume over time is shown for individual mice in each treatment group. Data are representative of two independent experiments.

[0122] [Figures 11A-11C]Figure 11 shows flow cytometry analysis of CD4+ TILs and tumor-draining lymph node T cells. BALB / C mice were inoculated with CT26 colorectal cancer cells. When tumors reached approximately 200 mm3 in size, mice were treated with isotype control, anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT antibodies for 7 days. Tumors and tumor-draining lymph nodes were collected. Data are representative of two independent experiments; n=5. Representative FACS plots gated on tumor-draining lymph node CD8+ T cells after in vitro stimulation; IFNγ-producing cells are boxed. IFNγ+ cells are quantified as a percentage of total CD8+ T cells. ***, P<0.001. CD8+ T cells are quantified as a percentage of total TILs. **, P=0.0065. Quantification of activated (CD44highCD62low) CD8+ T cells as a percentage of total CD8+ TILs. *, P=0.012. Quantification of CD8+ T cells as a percentage of total tumor-draining lymph node cells. Quantification of activated CD8+ T cells as a percentage of total CD8+ T cells in tumor-draining lymph nodes. *, P<0.05. Figure 11C shows quantification of CD4+ T cells as a percentage of total TILs. *, P=0.016. Figure 11A shows quantification of IFNγ+ cells as a percentage of CD4+ TILs after in vitro stimulation. Figure 11B shows quantification of IFNγ+ cells as a percentage of CD4+ T cells in tumor-draining lymph nodes after in vitro stimulation. Error bars indicate standard error of the mean. [Figures 11D-11F] Figure 11D shows quantification of activated CD4+ T cells as a percentage of total CD4+ TILs, Figure 11E shows quantification of CD4+ T cells as a percentage of total tumor-draining lymph node cells, and Figure 11F shows quantification of activated CD4+ T cells as a percentage of total CD4+ T cells in tumor-draining lymph nodes.

[0123] [Figure 12]Figure 12 shows further flow cytometry analysis of CD8+ TILs. BALB / C mice were inoculated with CT26 colorectal cancer cells and treated with isotype control, anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT antibodies as described in Figure 4. Tumors were harvested 7 days after treatment and analyzed by flow cytometry. Data are representative of two independent experiments; n=5. Figure 12A shows quantification of TNFα+ cells as a percentage of total CD8+ TILs. **, P<0.01. Figure 12B shows quantification of CD8+ TILs as a percentage of total TILs. **, P<0.01. Figure 12C shows quantification of activated (CD44highCD62Llow) CD8+ TILs as a percentage of total CD8+ TILs. *, P<0.05. Error bars indicate standard error of the mean.

[0124] [Figure 13A] Figure 13 shows flow cytometry analysis of tumor-draining lymph node-resident CD8+ T cells. BALB / C mice were inoculated with CT26 colorectal cancer cells and treated with isotype control, anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT antibodies as described in Figure 4. Tumor-draining lymph nodes were collected 7 days after treatment and analyzed by flow cytometry. Data are representative of two independent experiments; n = 5. Figure 13A shows a representative FACS plot gated on tumor-draining lymph node-resident CD8+ T cells after in vitro stimulation; IFNγ-producing cells are boxed. Quantification of IFNγ+ cells is as a percentage of total CD8+ T cells. ***, P < 0.001. [Figures 13B-13D] Figure 13B shows quantification of CD8+ T cells as a percentage of total cells in tumor-draining lymph nodes. Figure 13C shows quantification of activated (CD44highCD62Llow) CD8+ T cells as a percentage of total CD8+ T cells. *, P<0.05. Error bars indicate standard error of the mean. Figure 13D shows quantification of TNFα-producing cells as a percentage of total tumor-draining lymph node CD8+ T cells.

[0125] [Figure 14] Figure 14 shows co-expression of CD226 and TIGIT by tumor-infiltrating CD8+ T cells. C57BL6 / J mice were inoculated with MC38 colorectal cancer cells. Splenocytes and tumor-infiltrating lymphocytes (TILs) were analyzed approximately 14 days after inoculation, when tumors reached a size of approximately 200 mm3. Representative histograms of CD226 expression by splenic B cells (gray), splenic CD8+ T cells (blue), and TIGIT+ tumor-infiltrating CD8+ T cells (red). Data are representative of two independent experiments; n = 5.

[0126] [Figure 15] Figure 15 shows co-immunoprecipitation (co-IP) of CD226 and TIGIT on transfected cells. COS7 cells were co-transfected with expression plasmids containing cDNA for either TIGIT-HA (5 ng) or CD226-Flag (10 ng)-tagged proteins, or with a control plasmid (pRK). Following transfection, cells were washed and centrifuged to solubilize the cell pellet. The resulting supernatant was pre-cleared, centrifuged, and then equally divided into two tubes and immunoprecipitated with either anti-HA or anti-Flag using standard procedures. The immunoprecipitated proteins were subjected to SDS-PAGE and Western blot analysis. Western blots were probed with either anti-Flag-HRP or anti-HA-HRP.

[0127] [Figure 16] Figure 16 shows that TIGIT and CD226 interact in primary CD8+ T cells. MACS-enriched splenic C57BL6 / J CD8+ T cells were stimulated with plate-bound anti-CD3 and anti-CD28 antibodies and recombinant IL-2 for 48 hours and then solubilized. Cell lysates were immunoprecipitated with anti-TIGIT and probed with anti-CD226. Lanes: molecular weight ladder (1), input (2), co-immunoprecipitation flow-through (3), and co-immunoprecipitate. Arrows indicate the expected molecular weight of CD226.

[0128] [Figure 17] Figure 17 shows the detection of TIGIT / CD226 interaction by TR-FRET. Figure 17A shows the dissociation of Flag-ST-CD226 homodimers by HA-TIGIT. The FRET ratio between Flag-ST-CD226 was measured on COS-7 cells expressing a constant amount of Flag-ST-CD226 and increasing concentrations of HA-TIGIT. Figure 17B shows the FRET ratio between Flag-ST-CD226 recorded after 15 minutes of incubation with either PBS (white bars) or anti-TIGIT antibody (black bars). Figure 17C shows the binding of Flag-ST-CD226 to HA-TIGIT. The FRET intensity between Flag-ST-CD226 and HA-TIGIT in response to Flag-ST-CD226 expression was measured by anti-Flag ELISA on the same batch of transfected COS-7 cells. Figure 17D shows the FRET shift between Flag-ST-CD226 and HA-TIGIT after 15 minutes of incubation with PBS (white bars) or anti-TIGIT antibody (black bars). Data in A and C are representative of four independent experiments, each performed in triplicate. Data in B and D are representative of two independent experiments, each performed in triplicate.

[0129] [Figure 18] Figure 18 shows cell surface expression of Flag-ST-CD226 and HA-TIGIT. Anti-Flag and anti-HA ELISA on intact COS-7 cells expressing the indicated tagged constructs. Data are representative of three independent experiments, each performed in triplicate.

[0130] [Figures 19A-19B]Figure 19 shows that CD226 blockade reverses the enhanced antiviral T cell response induced by TIGIT / PD-L1 co-blockade. In Figures 19A-19D, C57BL6 / J mice were acutely depleted of CD4+ T cells and infected with clone 13 strain LCMV. Starting 28 days after infection, mice were treated with an isotype-matched control, anti-CD226, anti-PD-L1 + anti-TIGIT, or anti-PD-L1 + anti-TIGIT + anti-CD226 antibody. Splenocyte and liver viral titers were analyzed 42 days after infection. Figure 19A shows quantification of CD8+ T cells as a percentage of splenocytes. Figure 19B shows quantification of activated CD8+ T cells as a percentage of total CD8+ T cells. ***, P<0.001. [Figures 19C-19D] Figure 19C shows quantification of IFNγ-producing cells as a percentage of activated CD8+ T cells. ***, P<0.001. Figure 19D shows quantification of liver LCMV titers. ***, P<0.001. Error bars indicate standard error of the mean.

[0131] [Figure 20A] Figure 20 shows that TIGIT expression is elevated in human cancers and strongly correlates with CD8 and PD-1. Gene expression analysis of human cancers was performed as described in Example 11. Scatter plots show counts per gene data normalized by library size. Box and whisker plots show the variance-stabilized expression ratio of TIGIT and CD3e. Figure 20A shows the correlation between TIGIT and CD3e RNA expression in LUSC (gray) and normal lung (black). ρ=0.86. Quantification of the TIGIT / CD3e expression ratio is also shown. Increase in LUSC ratio=372%. ***, P=1.46×10-46. [Figure 20B] Figure 20B shows the correlation between TIGIT and CD3e RNA expression in COAD (gray) and normal colon (black). ρ=0.83. Quantification of the TIGIT / CD3e expression ratio is also shown. Increase in COAD ratio=116%. ***, P=3.66×10 [Figure 20C]Figure 20C shows the correlation between TIGIT and CD3e RNA expression in UCEC (gray) and normal endometrium (black). ρ=0.87. Quantification of the TIGIT / CD3e expression ratio is also shown. Increase in UCEC ratio=419%. ***, P=7.41×10-5. [Figure 20D] Figure 20D shows the correlation between TIGIT and CD3e RNA expression in BRCA (gray) and normal breast (black). ρ=0.82. Quantification of the TIGIT / CD3e expression ratio is also shown. Increase in BRCA ratio=313%. ***, P=4.6×10-44. [Figure 20E] Figure 20E shows the correlation between TIGIT and CD3e RNA expression in renal clear cell carcinoma (gray) and normal kidney (black). ρ=0.94. Quantification of the TIGIT / CD3e expression ratio is also shown. [Figure 20F] Figure 20F shows the correlation between TIGIT and CD8A (left) or TIGIT and CD4 (right) in lung squamous cell carcinoma (gray) and normal lung (black), ρ = 0.77 and 0.48, respectively. [Figures 20G-20H] Figure 20G shows the correlation between TIGIT and PD-1 (Pdcd1) in lung squamous cell carcinoma (gray) and normal lung (black). ρ=0.82. Figure 20H shows the correlation between TIGIT and CD226 in lung squamous cell carcinoma (red) and normal lung (black). ρ=0.64.

[0132] [Figure 21] Figure 21 shows an analysis of T cell-associated gene expression in lung squamous cell carcinoma (LUSC). Gene expression in LUSC and normal tissue samples was analyzed as described in Example 11, and a heatmap of genes that best correlated with the gene signature in LUSC samples was generated. Genes and samples were both clustered using hierarchical clustering using Ward linkage on a Euclidean distance matrix on centered and scaled expression data.

[0133] [Figures 22A-22B]Figure 22 shows that TIGIT and PD-1 are coordinately expressed by human and mouse tumor-infiltrating lymphocytes. Figures 22A-22C show analyses of lymphocytes from freshly resected human NSCLC tumors, tumor-matched peripheral blood, and peripheral blood from normal donors. Data are representative of tumors analyzed twice independently. Figure 22A shows a representative FACS plot depicting TIGIT expression by peripheral and tumor-infiltrating CD8+ T cells, with TIGIT+ cells boxed. Figure 22B shows a representative FACS plot depicting TIGIT expression by peripheral and tumor-infiltrating CD4+ T cells, with TIGIT+ cells boxed. [Figures 22C-22E] Figure 22C shows flow cytometry histograms depicting TIGIT expression by PD-1 high (red) and PD-1 low (blue) NSCLC-infiltrating CD8+ (left) and CD4+ (right) T cells. In Figures 22D-22G, BALB / C mice were inoculated with syngeneic CT26 colorectal cancer cells. Splenocytes and tumor-infiltrating lymphocytes (TILs) were analyzed 14 days after inoculation, when tumors had reached a size of approximately 200 mm. Data are representative of two independent experiments; n = 5-6. Figure 22D shows a representative FACS plot of TIGIT expression by tumor-infiltrating CD8+ T cells; TIGIT+ cells are boxed. Figure 22E shows a representative FACS plot of TIGIT expression by tumor-infiltrating CD4+ T cells; TIGIT+ cells are boxed. Quantification of the frequency of TIGIT+ T cells is as a percentage of total T cells. *, P = 0.0134. ***, P < 0.0001. [Figures 22F-22G] Figure 22F shows flow cytometry histograms representing TIGIT expression by PD-1 high and PD-1 low tumor-infiltrating CD8+ T cells and by splenic CD8+ T cells. Quantification of TIGIT MFI is also shown. **, P=0.0023. Figure 22G shows flow cytometry histograms representing TIGIT expression by PD-1 high and PD-1 low tumor-infiltrating CD4+ T cells and by splenic CD4+ T cells. Quantification of TIGIT MFI is also shown. ***, P=0.0002. Error bars indicate standard error of the mean.

[0134] [Figures 23A-23C] Figure 23 shows the characterization of TIGIT expression by human tumor-infiltrating T cells. Figures 23A-23B show FACS plots showing TIGIT expression by NSCLC tumor-infiltrating CD8+ and CD4+ T cells (Figure 23A) and by donor-matched PBMC CD8+ and CD4+ T cells (Figure 23B), with TIGIT+ cells boxed. Figure 23C shows FACS plots showing TIGIT expression by CRC tumor-infiltrating CD8+ and CD4+ T cells (Figure 23C), with TIGIT+ cells boxed. [Figure 23D] FACS plots showing TIGIT expression by donor-matched PBMC CD8+ and CD4+ T cells (FIG. 23D) are shown, with TIGIT+ cells boxed.

[0135] [Figure 24] Figure 24 shows that the TIGIT:CD226 interaction is not driven by PVR TIGIT:CD226 and TIGIT Q56R:CD226 interactions detected by TR-FRET, and the FRET ratios between Flag-ST-CD226 and HA-TIGIT or HA-TIGIT Q56R indicate that WT and Q56R TIGIT bind CD226 with the same potency. Data are representative of three independent experiments performed in triplicate.

[0136] [Figure 25A] Figure 25 shows the effect of TIGIT / PD-L1 antibody co-blockade in MC38 tumor-bearing mice. In Figures 25A-25C, MC38 tumor-bearing mice were generated as described above and treated for 3 weeks with blocking antibodies against PD-L1 (red), TIGIT (blue), TIGIT and PD-L1 (purple), or an isotype-matched control antibody (black). N = 10 (control, anti-PD-L1 alone, anti-TIGIT alone) or 20 (anti-TIGIT + anti-PD-L1). Figure 25A shows the median (left) and individual values ​​(right) of MC38 tumor volume over time. [Figures 25B-25C] Figure 25B shows MC38 tumor volume 14 days after antibody treatment. ***, P=0.0005. **, P=0.0093. *, P=0.0433. Figure 25C shows mouse survival over time. Error bars indicate standard error of the mean.

[0137] [Figures 26A-26C] Figure 26 shows further characterization of TIGIT expression by mouse tumor-infiltrating T cells. Figure 26A shows splenic C57BL6 / J CD8+ T cells enriched by MACS and cultured with plate-coated anti-CD3 and anti-CD28 agonist antibodies. Representative histograms of TIGIT (red) and isotype-matched control (gray fill) staining over time. Quantification of TIGIT MFI. ***, P<0.001. Stimulated cells inducibly expressed PD-1 and constitutively expressed CD226 (data not shown). Data are representative of two independent experiments; n=5. In Figures 26B-26E, wild-type C57BL6 / J mice were subcutaneously inoculated with syngeneic MC38 colorectal cancer cells. Tumors were allowed to grow without intervention until they reached a size of 150-200 mm3. Data are representative of two independent experiments; n=5. Figure 26B shows a representative FACS plot of tumor-infiltrating CD8+ T cells, with TIGIT+ cells boxed. Quantification of the frequency of TIGIT+ cells as a percentage of total tumor-infiltrating or splenic CD8+ T cells. ***, P<0.0001. Figure 26C shows a representative FACS plot of tumor-infiltrating CD4+ T cells, with TIGIT+ cells boxed. Quantification of the frequency of TIGIT+ cells as a percentage of total tumor-infiltrating or splenic CD4+ T cells. ***, P<0.0001. [Figure 26D-26E]Figure 26D shows representative histograms of TIGIT expression by PD-1 high and PD-1 low tumor-infiltrating CD8+ T cells (red and blue, respectively) and by splenic CD8+ T cells (gray). Quantification of TIGIT MFI. ***, P<0.0001. Figure 26E shows representative histograms of TIGIT expression by PD-1 high and PD-1 low tumor-infiltrating CD4+ T cells and by splenic CD4+ T cells. Quantification of TIGIT MFI. *, P=0.0136. **, P=0.0029. Error bars indicate standard error of the mean.

[0138] [Figure 27] Figure 27 shows that tumor-infiltrating CD8+ and CD4+ T cells maintain high levels of CD226 expression. Wild-type BALB / c mice were inoculated with the CT26 tumor cells described herein. After tumors reached approximately 150-200 mm in size, tumors and spleens were analyzed by flow cytometry. Figure 27A shows quantification of CD226+ CD8+ T cells, CD4+ T cells, and non-T cells as percentages of total CD8+ T cells, CD4+ T cells, and non-T cells, respectively. Figure 27B shows representative histograms of CD226 expression in tumors and spleens. Data are representative of two independent experiments; n = 5. Error bars indicate standard error of the mean.

[0139] [Figure 28A] Figure 28 shows that TIGIT suppression of CD8+ T cell responses is dependent on CD226. BALB / C mice were inoculated subcutaneously with CT26 colorectal cancer cells in the right thoracic flank. When tumors reached approximately 200 mm3 in size, mice were treated for 3 weeks with isotype control (black), anti-CD226 (orange), anti-PD-L1 (red), anti-TIGIT + anti-PD-L1 (purple), or anti-TIGIT + anti-PD-L1 + anti-CD226 (green) antibodies. Data are representative of one experiment; n = 10 (AB) or 5 (CF). Figure 28A shows the median (left) and individual (right) values ​​of CT26 tumor volume over time. [Fig. 28B-28F]Figure 28B shows mouse survival over time. In Figures 28C-28F, tumor-infiltrating lymphocytes and tumor-draining lymph node-resident lymphocytes were assessed by flow cytometry 7 days after treatment. Figure 28C shows quantification of IFNγ-producing CD8+ TILs as a percentage of total CD8+ TILs after in vitro stimulation. **, P<0.01. Figure 28D shows quantification of IFNγ-producing cells as a percentage of total CD8+ T cells after in vitro stimulation. *, P<0.05. Figure 28E shows quantification of CD8+ TILs as a percentage of total TILs. **, P<0.01. Figure 28F shows quantification of CD8+ T cells as a percentage of total tumor-draining lymph node-resident lymphocytes. Error bars indicate standard error of the mean.

[0140] [Figures 29A-29D]Figure 29 shows that TIGIT impairs CD226 function by interfering with CD226 homodimerization. Figure 29A shows CD8+ T cells were enriched by MACS from TIGITfl / flCD4cre (CKO) and TIGITfl / flCD4wt (WT) littermates and stimulated in the presence of anti-CD226 or isotype-matched control antibodies, as indicated. H3-thymidine incorporation is shown as the ratio of cells cultured with anti-CD3 + PVR-Fc to cells cultured with anti-CD3 alone. **, P=0.0061. ***, P<0.0001. Data are representative of two independent experiments; n=5. Figure 29B shows wild-type C57BL6 / J CD8+ T cells were enriched by MACS and stimulated in the presence of anti-TIGIT, anti-CD226, and / or isotype-matched control antibodies, as indicated. H3-thymidine incorporation is shown as the ratio of cells cultured with anti-CD3 + PVR-Fc to cells cultured with anti-CD3 alone. ***, P<0.001 in a paired t-test. Figure 29C shows primary human CD8+ T cells were MACS-enriched from blood and stimulated with suboptimal levels of plate-bound anti-CD3 in the presence or absence of human recombinant PVR-Fc. Anti-TIGIT antibody or an isotype-matched control antibody was added as indicated. Quantification of H-thymidine incorporation. **, P=0.0071 and 0.0014, respectively. Figure 29D shows CHO cells were transiently transfected with increasing concentrations of acceptor and donor FLAG-ST-CD226 as indicated. Quantification of FRET intensity relative to donor emission. Data are representative of three independent experiments; n=3. [Figures 29E-29H]In Figures 29E-29F, CHO cells were transiently transfected with FLAG-ST-CD226 and increasing concentrations of HA-TIGIT, as indicated. Data are representative of two or more independent experiments; n = 4. Data are normalized to the maximum signal. Figure 29E shows quantification of the CD226:CD226 FRET ratio (FRET ratio 1). Figure 29F shows quantification of the TIGIT:CD226 FRET ratio (FRET ratio 2). Figure 29G shows anti-FLAG (left) and anti-HA (right) immunoblots performed with either anti-FLAG or anti-HA immunoprecipitations prepared from COS-7 cells transfected with either the empty pRK vector or a combination of Flag-CD226 and HA-TIGIT. Data are representative of two independent experiments. Figure 29H shows quantification of the TIGIT:CD226 FRET ratio after incubation with PBS (white) or anti-TIGIT antibody (red). ***, P<0.001. Data are representative of four independent experiments; n=3. Error bars indicate the standard error of the mean.

[0141] [Figure 30] Figure 30 shows that primary human T cells were MACS-enriched from blood and stimulated with anti-CD3 and anti-CD28. TIGIT+ and TIGIT- cells were sorted, rested, restimulated, and labeled for FRET with the indicated antibodies. Data are representative of two independent experiments. ***, P<0.001. Error bars indicate standard error of the mean.

[0142] [Figure 31]Figure 31 shows that TIGIT and PD-1 co-blockade does not restore the effector function of exhausted CD4+ T cells during chronic viral infection. Figure 31A shows quantification of CD8+ T cells as a percentage of total splenocytes. Figure 31B shows gp33 pentamer+ cells as a percentage of total splenic CD8+ T cells. **, P=0.0040. Figure 31C shows a representative FACS plot gated on gp33 pentamer+ CD8+ T cells after in vitro stimulation, with IFN-gamma+ cells boxed. Quantification of IFN-γ-producing cells as a percentage of total gp33 pentamer+ CD8+ T cells. *, P=0.0319. **, P=0.0030. Error bars indicate standard error of the mean.

[0143] [Figure 32A] Figure 32 shows that the effect of TIGIT / PD-L1 co-blockade is dependent on CD8+ T cells. In Figures 32A-32B, wild-type BALB / c mice were inoculated with CT26 tumors as described in Figure 7. When tumors reached a size of 100-150 mm3, the mice were transiently depleted of CD8+ T cells and treated with anti-TIGIT + anti-PD-L1. Data are representative of one experiment; n=10 / group. Figure 32A shows the median (left) and individual values ​​(right) of CT26 tumor volume over time. [Fig. 32B-32C] Figure 32B shows quantification of CT26 tumor volume 17 days after treatment initiation. ***, P=0.0004. In Figure 32C, wild-type BALB / c mice were inoculated with CT26 tumors and treated with anti-TIGIT + anti-PD-L1, followed by re-challenge with CT26 tumors that were transiently depleted of CD8+ T cells at the time of re-challenge. Data are representative of two experiments; n=5. Figure 32C shows median (left) and individual values ​​(right) of CT26 tumor volume over time. Error bars indicate standard error of the mean.

[0144] [Figure 33]Figure 33 shows that PVR expression on tumor cells is not required for the TIGIT / PD-L1 co-blockade effect. Wild-type BALB / c mice were inoculated with wild-type or PVR-deficient (PVR.KO) tumors as described. When tumors reached a size of 150-200 mm, mice were treated with anti-TIGIT + anti-PD-L1 or isotype-matched control antibodies. Data are representative of one experiment; n=10 / group. Figure 33 shows the median (left) and individual values ​​(right) of CT26 tumor volume over time.

[0145] [Figure 34] Figure 34 shows the effect of TIGIT / PD-L1 antibody co-blockade in EMT6 tumor-bearing mice. EMT6 tumor-bearing mice were generated as described above and treated for 3 weeks with blocking antibodies against PD-L1 (red), TIGIT (blue), TIGIT and PD-L1 (purple), or an isotype-matched control antibody (black). N=10 (control, anti-PD-L1 alone, anti-TIGIT alone) or 20 (anti-TIGIT + anti-PD-L1). Figure 34 shows the median (left) and individual values ​​(right) of EMT6 tumor volume over time.

[0146] [Figure 35]Figure 35 shows that TIGIT regulates tumor-infiltrating CD8+ T cell effector function. BALB / C mice were inoculated subcutaneously with CT26 colorectal cancer cells in the right thoracic flank and treated with anti-PD-L1, anti-TIGIT, or anti-PD-L1 + anti-TIGIT as described in Figure 7. Tumor-draining lymph node (dLN)-resident and tumor-infiltrating T cells were analyzed by flow cytometry 7 days after the start of treatment. Data are representative of two independent experiments; n = 5. Figure 35A shows quantification of IFNγ / TNFα dual-producing dLN-resident CD8+ and CD4+ T cells as a percentage of total dLN-resident CD8+ and CD4+ T cells, respectively. Dual cytokine production by unstimulated T cells is also shown. **, P = 0.002, 0.003, and 0.001, respectively. Figure 35B shows quantification of IFNγ / TNFα dual-producing tumor-infiltrating CD8+ and CD4+ T cells as a percentage of total tumor-infiltrating CD8+ and CD4+ T cells, respectively. Dual cytokine production by unstimulated T cells is also shown. ***, P<0.0001. Error bars indicate standard error of the mean.

[0147] [Figure 36] Figure 36 shows analysis of lymphocytes from resected human NSCLC tumors, tumor-matched peripheral blood, and normal donor peripheral blood. Data are pooled from three independently acquired sample sets. Figure 36A shows quantification of TIGIT+ cells as a percentage of total CD8+ T cells. *, P<0.05. Figure 36B shows quantification of TIGIT+ cells as a percentage of total CD4+ T cells.

[0148] [Figure 37]Figure 37 shows the characterization of TIGIT expression in human tumors. Figure 37A shows representative flow cytometry histograms of TIGIT expression by NSCLC tumor-resident lymphocytes (red, CD45+FSClow), myeloid cells (blue, CD45+FSChigh), and non-hematopoietic cells (green, CD45-) relative to subset-matched isotype staining (gray). Figure 37B shows the gating strategy for PD-1high and PD-1low NSCLC tumor-infiltrating CD8+ and CD4+ T cells. DETAILED DESCRIPTION OF THE INVENTION

[0149] I. General techniques The techniques and procedures described or referenced herein are generally well understood by those skilled in the art and may be found, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed. (2001), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., (2003)); the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames, and G.R. Taylor, eds. (1995)), Harlow and Lane, eds. (1988) Antibodies, A Laboratory Manual, and Animal Cell Culture (R.I. Freshney, ed. (1987)); Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (JP Mather and PE Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, JB Griffiths, and DG Newell, eds., 1993-8) J. Wiley and Sons; Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JMMiller and MP Calos, eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (JE Coligan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty, ed., IRL Press, 1988-1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995); and the widely used methods described in Cancer: Principles and Practice of Oncology (eds. VT DeVita et al., JB Lippincott Company, 1993).

[0150] II. Definition The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners, with the result that T cell function (e.g., proliferation, cytokine production, target cell killing) is restored or enhanced, eliminating T cell dysfunction resulting from signaling along the PD-1 signaling axis. As used herein, PD-1 axis binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0151] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates, or interferes with signal transduction resulting from the interaction of PD-1 with one or more of its binding partners, e.g., PD-L1, PD-L2. In some embodiments, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its binding partners. In particular aspects, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or interfere with signal transduction resulting from the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, the PD-1 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-1, such that dysfunction of dysfunctional T cells is reduced (e.g., effector responses to antigen recognition are enhanced). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a specific embodiment, the PD-1 binding antagonist is MDX-1106, as described herein. In another specific embodiment, the PD-1 binding antagonist is Merck 3745, as described herein. In another specific embodiment, the PD-1 binding antagonist is CT-011, as described herein. In another specific embodiment, the PD-1 binding antagonist is AMP-224, as described herein.

[0152] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates, or interferes with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, e.g., PD-1 and B7-1. In some embodiments, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partners. In particular aspects, PD-L1 binding antagonists inhibit the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or interfere with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, e.g., PD-1 and B7-1. In one embodiment, the PD-L1 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L1, such that the dysfunctional T cells are less dysfunctional (e.g., enhance their effector response to antigen recognition). In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In a particular embodiment, the anti-PD-L1 antibody is YW243.55.S70 described herein. In another particular embodiment, the anti-PD-L1 antibody is MDX-1105 described herein. In yet another particular embodiment, the anti-PD-L1 antibody is MPDL3280A described herein. In another particular embodiment, the anti-PD-L1 antibody is MEDI4736 described herein.

[0153] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates, or interferes with signaling resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In some embodiments, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to its binding partners. In a particular aspect, a PD-L2 binding antagonist inhibits the binding of PD-L2 to PD-1. In some embodiments, PD-L2 antagonists include anti-PD-L2 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or interfere with signaling resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In one embodiment, the PD-L2 binding antagonist reduces the negative costimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L2, such that the dysfunctional T cells are less dysfunctional (e.g., enhance the effector response to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0154] The term "aptamer" refers to a nucleic acid molecule capable of binding to a target molecule, such as a polypeptide. For example, the aptamer of the present invention can specifically bind to a TIGIT polypeptide or a molecule in a signaling pathway that regulates the expression of TIGIT. The generation and therapeutic use of aptamers is well established in the art. See, for example, U.S. Pat. No. 5,475,096 and the therapeutic efficacy of Macugen® (Eyetech, New York) for the treatment of age-related macular degeneration.

[0155] The term "antagonist" is used in the broadest sense and includes any molecule that partially or fully blocks, inhibits, or neutralizes a biological activity of a native polypeptide disclosed herein. Similarly, the term "agonist" is used in the broadest sense and includes any molecule that mimics a biological activity of a native polypeptide disclosed herein. Suitable agonist or antagonist molecules include, among others, agonist or antagonist antibodies or antibody fragments, fragments or amino acid sequence variants of native polypeptides, peptides, antisense oligonucleotides, small organic molecules, and the like. Methods for identifying agonists or antagonists of a polypeptide may involve contacting the polypeptide with a candidate agonist or antagonist molecule and measuring a detectable change in one or more biological activities normally associated with the polypeptide.

[0156] The terms "TIGIT antagonist" and "antagonist of TIGIT activity or expression" are used interchangeably and refer to a compound that interferes with the normal function of TIGIT by reducing the transcription or translation of TIGIT-encoding nucleic acid, or by inhibiting or blocking TIGIT polypeptide activity, or both. Examples of TIGIT antagonists include, but are not limited to, antisense polynucleotides, interfering RNAs, catalytic RNAs, RNA-DNA chimeras, TIGIT-specific aptamers, anti-TIGIT antibodies, TIGIT-binding fragments of anti-TIGIT antibodies, TIGIT-binding small molecules, TIGIT-binding peptides, and other polypeptides that specifically bind to TIGIT (including, but not limited to, TIGIT-binding fragments of one or more TIGIT ligands, optionally fused with one or more other domains) such that interaction of the TIGIT antagonist with TIGIT results in the reduction or cessation of TIGIT activity or expression. It will be understood by those skilled in the art that in some cases, a TIGIT antagonist may antagonize one TIGIT activity without affecting other TIGIT activities. For example, a desirable TIGIT antagonist for use in some of the methods herein is one that antagonizes TIGIT activity in response to one of PVR interaction, PVRL3 interaction, or PVRL2 interaction, for example, without affecting or minimally affecting any of the other TIGIT interactions.

[0157] The terms "PVR antagonist" and "antagonist of PVR activity or expression" are used interchangeably and refer to compounds that interfere with the normal function of PVR by reducing the transcription or translation of PVR-encoding nucleic acid, inhibiting or blocking PVR polypeptide activity, or both. Examples of PVR antagonists include, but are not limited to, antisense polynucleotides, interfering RNA, catalytic RNA, RNA-DNA chimeras, PVR-specific aptamers, anti-PVR antibodies, PVR-binding fragments of anti-PVR antibodies, PVR-binding small molecules, PVR-binding peptides, and other polypeptides (including, but not limited to, PVR-binding fragments of one or more PVR ligands, optionally fused to one or more other domains) that specifically bind to PVR such that interaction of the PVR antagonist with PVR results in a reduction or cessation of PVR activity or expression. Those skilled in the art will understand that, in some cases, a PVR antagonist may antagonize one PVR activity without affecting other PVR activities. For example, desirable PVR antagonists for use in certain of the methods herein are those that antagonize PVR activity in response to TIGIT interaction without affecting PVR-CD96 and / or PVR-CD226 interaction.

[0158] The term "dysfunction" in the context of immune dysfunction refers to a state of diminished immune responsiveness to antigenic stimulation. The term includes the common elements of both exhaustion and / or anergy, in which antigen recognition can occur but the subsequent immune response is ineffective in controlling infection or tumor growth.

[0159] As used herein, the term "dysfunctional" includes refractoriness or unresponsiveness to antigen recognition, particularly an impaired ability to translate antigen recognition into downstream T cell effector functions such as proliferation, cytokine production (e.g., IL-2) and / or target cell killing.

[0160] The term "anergy" refers to a state of unresponsiveness to antigenic stimulation resulting from defective or insufficient signals delivered via the T cell receptor (e.g., intracellular Ca in the absence of ras activation). +2 T cell anergy can also occur upon stimulation with antigen in the absence of costimulation, rendering the cells refractory to subsequent activation by antigen even in the context of costimulation. The unresponsive state can often be reversed by the presence of interleukin-2. Anergic T cells do not undergo clonal expansion and / or acquire effector function.

[0161] The term "exhaustion" refers to T cell exhaustion, a state of T cell dysfunction resulting from persistent TCR signaling, which occurs during many chronic infections and cancers. It is distinct from anergy in that it results from persistent signaling rather than through defective or insufficient signaling. It is defined by poor effector function, persistent expression of inhibitory receptors, and a transcriptional state that differs from that of functional effector or memory T cells. Exhaustion prevents optimal control of infections and tumors. Exhaustion can result from both extrinsic negative regulatory pathways (e.g., immunomodulatory cytokines) and cell-intrinsic negative regulatory (costimulatory) pathways (e.g., PD-1, B7-H3, B7-H4).

[0162] "Enhancing T cell function" means inducing, encouraging, or stimulating T cells to have sustained or amplified biological function, or regenerating or reactivating exhausted or inactive T cells. Examples of enhancing T cell function include increasing the number of CD8 T cells relative to their levels before the intervention. + These include increased secretion of gamma interferon from T cells, increased proliferation, and increased antigen responsiveness (e.g., viral, pathogen, or tumor clearance). In one embodiment, the level of enhancement is at least 50%, or 60%, 70%, 80%, 90%, 100%, 120%, 150%, or 200%. Methods for measuring this enhancement are known to those skilled in the art.

[0163] A "T cell dysfunction disorder" is a disease or condition of T cells characterized by decreased responsiveness to antigenic stimulation. In certain embodiments, the T cell dysfunction disorder is one specifically associated with increased inappropriate signaling through PD-1. In another embodiment, the T cell dysfunction disorder is one in which T cells are anergic or have a reduced ability to secrete cytokines, proliferate, or carry out cytolytic activity. In certain aspects, the reduced responsiveness results in ineffective control of immunogen-expressing pathogens or tumors. Examples of T cell dysfunction disorders characterized by T cell dysfunction include unresolved acute infections, chronic infections, and tumor immunity.

[0164] "Tumor immunity" refers to the process by which tumors evade immune recognition and clearance. Thus, as a therapeutic concept, tumor immunity is "treated" when such evasion is attenuated and the tumor is recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor clearance.

[0165] "Immunogenicity" refers to the ability of a particular substance to induce an immune response. Tumors are immunogenic, and enhancing tumor immunogenicity aids in the clearance of tumor cells by the immune response. Examples of enhancing tumor immunogenicity include, but are not limited to, treatment with PD-1 axis binding antagonists (e.g., anti-PD-L1 antibodies) and TIGIT inhibitors (e.g., anti-TIGIT antibodies).

[0166] "Dured response" refers to a sustained effect on tumor growth reduction after treatment is discontinued. For example, tumor size may remain the same or smaller compared to the size at the beginning of the administration phase. In some embodiments, the sustained response is at least as long as the treatment period, or at least 1.5x, 2.0x, 2.5x, or 3.0x the length of the treatment period.

[0167] The term "antibody" includes monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region), antibody compositions with polyepitopic specificity, multispecific antibodies (e.g., bispecific antibodies, diabodies, and single-chain molecules), and antibody fragments (e.g., Fab, F(ab'), and Fv). The term "immunoglobulin" (Ig) is used interchangeably with "antibody" herein.

[0168] The basic four-chain antibody unit is a heterotetrameric glycoprotein consisting of two identical light (L) chains and two identical heavy (H) chains. IgM antibodies consist of five basic heterotetrameric units together with an additional polypeptide called the J chain and have 10 antigen-binding sites, while IgA antibodies contain 2-5 basic four-chain units that can polymerize to form multivalent assemblies with the J chain. In the case of IgG, the four-chain unit is generally about 150,000 daltons. Each L chain is linked to an H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain contains a variable domain (V) at its N-terminus. H ), followed by three constant domains (C H ) and four C for μ and ε isotypes H Each L chain has a variable domain (V L ) followed by a constant domain at the other end. L is V H are aligned in C L is the first constant domain of the heavy chain (C H 1) are aligned. Specific amino acid residues are thought to form an interface between the light- and heavy-chain variable domains. V H and V LThese antibodies combine in pairs to form a single antigen-binding site. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th ed., Daniel P. Stites, Abba I. Terr, and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, p. 71, Chapter 6. Light chains from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequence of their constant domains. Depending on the amino acid sequence of the constant domain (CH) of their heavy chains, immunoglobulins can be assigned to various classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated α, δ, ε, γ, and μ, respectively. The gamma and alpha classes are further divided into subclasses based on relatively minor differences in CH sequence and function; for example, humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0169] By "variable region" or "variable domain" of an antibody is meant the amino-terminal domain of the heavy or light chain of the antibody. The heavy and light chain variable domains may also be referred to as "VH" and "VL," respectively. These domains are generally the most variable parts of an antibody (relative to other antibodies of the same class) and contain the antigen-binding site.

[0170] The term "variable" refers to the fact that certain segments of the variable domains differ extensively in sequence among antibodies. The V domain mediates antigen binding and determines the specificity of a particular antibody for its particular antigen. However, variability is not uniformly distributed throughout the entire span of the variable domains. Instead, it is concentrated in three segments called hypervariable regions (HVRs) in both the light- and heavy-chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs). Native heavy- and light-chain variable domains each contain four FR regions that primarily adopt a β-sheet configuration, connected by three HVRs that form loops that connect, and in some cases form part of, the β-sheet structure. The HVRs of each chain are held in close proximity by the FR regions and, together with HVRs from other chains, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Immunological Interest, 5th ed., National Institutes of Health, Bethesda, MD. (1991)). The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity.

[0171] The term "monoclonal antibody," as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies within the population are identical except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation), which may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they are synthesized by a hybridoma culture, uncontaminated by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in the present invention can be produced using hybridoma methods (e.g., Kohler and Milstein, Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14(3):253-260 (1995); Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al., Monoclonal Antibodies and T-Cell Hybridomas, 563-681 (Elsevier, NY, 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), phage display techniques (e.g., Clackson et al., Nature, 352:624-628 (1991); Marks et al., J. Mol. Biol., 222:581-597 (1992); Sidhu et al., J. Mol. Biol., 338(2):299-310(2004); Lee et al., J. Mol. Biol., 340(5):1073-1093(2004); Fellouse, Proc. Natl. Acad. Sci. USA, 101(34):12467-12472(2004); and Lee et al., J. Immunol.Methods, 284(1-2):119-132 (2004), and techniques for producing human or human-like antibodies in animals having part or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immunol., 7:33 (1993); U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al., Bio / Technology, 10:779-783 (1992); Lonberg et al., Nature, 368:856-859 (1994); Morrison, Nature, 368:812-813 (1994); Fishwild et al., Nature Biotechnol., 14:845-851 (1996); Neuberger, Nature Biotechnol., 14:826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol., These can be produced by a variety of techniques, including those described in U.S. Pat. No. 13:65-93 (1995).

[0172] The term "naked antibody" means an antibody that is not conjugated to a cytotoxic moiety or radiolabel.

[0173] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably and refer to an antibody in its substantially intact form, as opposed to an antibody fragment. In particular, whole antibodies include those having heavy and light chains, including an Fc region. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some cases, an intact antibody may have one or more effector functions.

[0174] An "antibody fragment" comprises a portion of an intact antibody, preferably the antigen-binding and / or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (U.S. Pat. No. 5,641,870, Example 2; Zapata et al., Protein Eng. 8(10): 1057-1062

[1995] ); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Papain digestion of an antibody produces two identical antigen-binding fragments called "Fab" fragments and a residual "Fc" fragment, a designation reflecting its ability to crystallize readily. Fab fragments contain the heavy chains (V H ) variable region domain of one heavy chain, and the first constant domain of one heavy chain (C H 1) and an entire L chain. Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. Pepsin treatment of an antibody yields a single large F(ab')2 fragment which roughly corresponds to two disulfide-linked Fab fragments with different antigen-binding activities and is still capable of cross-linking antigen. The Fab' fragment contains one or more cysteines from the antibody hinge region. H F(ab')2 antibody fragments differ from Fab fragments by having a few additional residues at the carboxy terminus of one domain. Fab'-SH is the designation used here for Fab' in which the cysteine ​​residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments are produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical linkages of antibody fragments are also known.

[0175] The Fc fragment contains the carboxy-terminal portions of both heavy chains held together by disulfides. The effector functions of an antibody are determined by sequences in the Fc region, which is also recognized by Fc receptors (FcRs) found on certain cells.

[0176] An "Fv" is the minimum antibody fragment containing a complete antigen-recognition and binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. The folding of these two domains results in six hypervariable loops (three loops each from the H and L chain) that contribute amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although with a lower affinity than the entire binding site.

[0177] "Single-chain Fv," also abbreviated as "sFv" or "scFv," refers to VFvs linked to a single polypeptide chain. H and V L Preferably, the sFv polypeptide contains a polypeptide linker which enables the sFv to form the desired structure for antigen binding. H and V L For a review of sFvs, see Pluckthun, The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore (eds.), Springer-Verlag, New York, pp. 269-315 (1994).

[0178] "Functional fragments" of antibodies of the present invention include portions of the intact antibody, generally including the antigen-binding or variable region of the intact antibody, or the Fc region of the antibody that retains FcR binding ability or has modified FcR binding ability. Examples of antibody fragments include linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0179] The term "diabody" refers to a diabody that combines V domains such that inter-chain, but not intra-chain, pairing of the V domains is achieved, thus resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. H and V LIt refers to small antibody fragments prepared by constructing sFv fragments (see previous paragraph) with a short linker (about 5-10 residues) between the domains. Bispecific diabodies are small antibody fragments prepared by constructing sFv fragments (see previous paragraph) with a short linker (about 5-10 residues) between the domains. H and V L Diabodies are heterodimers of two "crossover" sFv fragments in which the domains are present on different polypeptide chains. Diabodies are described in further detail in, for example, EP 404097; WO 93 / 11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993).

[0180] The monoclonal antibodies herein specifically include "chimeric" antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical to or homologous to the corresponding sequence of an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical to or homologous to the corresponding sequence of an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they possess the desired biological activity (U.S. Pat. No. 4,816,567, and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). Chimeric antibodies of interest herein include PRIMATIZED® antibodies, in which the antigen-binding region of the antibody is derived from an antibody produced, for example, by immunizing macaque monkeys with the antibody of interest. As used herein, "humanized antibody" refers to a subset of "chimeric antibody."

[0181] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from an HVR (defined below) of the recipient are replaced by residues from an HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and / or capacity. In some instances, framework ("FR") residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in either the recipient antibody or the donor antibody. These modifications may be made to further refine antibody performance, e.g., binding affinity. Generally, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or almost all of the hypervariable loops correspond to those of a non-human immunoglobulin sequence, and all or almost all of the FR regions are those of a human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions that improve antibody performance, e.g., binding affinity, isomerization, immunogenicity, etc. The number of these amino acid substitutions in the FRs will typically be no more than six in the H chain and no more than three in the L chain. The humanized antibody optionally will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see, e.g., Jones et al., Nature 321, 522-525 (1986); Reichmann et al., Nature 332, 323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2, 593-596 (1992). See also, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol., 1:105-115 (1998); Harris, Biochem. Soc. Transactions, 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech., 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409.

[0182] A "human antibody" is an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and / or an antibody produced using any of the techniques for producing human antibodies disclosed herein. This definition of human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also useful for preparing human monoclonal antibodies are the methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering antigen to transgenic animals, e.g., immunized xenomouse, which have been modified to produce such antibodies in response to antigen challenge, but which have been disabled at their endogenous locus (e.g., XENOMOUSE TM (See U.S. Patent Nos. 6,075,181 and 6,150,584 for related technology.) Also, see, e.g., Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006), for human antibodies produced via human B cell hybridoma technology.

[0183] As used herein, the terms "hypervariable region," "HVR," or "HV" refer to regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops. Antibodies generally contain six HVRs: three in the VH (H1, H2, and H3) and three in the VL (L1, L2, and L3). In natural antibodies, H3 and L3 represent the greatest diversity of the six HVRs, and H3 in particular is thought to play a unique role in conferring fine specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, NJ, 2003). Indeed, naturally occurring camelid antibodies consisting only of heavy chains are functional and stable in the absence of light chains. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0184] Numerous HVR delineations are in use and are encompassed herein. Kabat complementarity-determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat, S., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia instead refers to the location of structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). AbM HVRs represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. "Contact" HVRs are based on analysis of available complex crystal structures. The residues in each of these HVRs are listed below. Loop Kabat AbM Chothia Contact L1 L24-L34 L24-L34 L26-L32 L30-L36 L2 L50-L56 L50-L56 L50-L52 L46-L55 L3 L89-L97 L89-L97 L91-L96 L89-L96 H1 H31-H35B H26-H35B H26-H32 H30-H35B (Kabat numbering) H1 H31-H35 H26-H35 H26-H32 H30-H35 (Chothia numbering) H2 H50-H65 H50-H58 H53-H55 H47-H58 H3 H95-H102 H95-H102 H96-H101 H93-H101

[0185] HVRs may include "extended HVRs" as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, and 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH. For each of these definitions, the variable domain residues are numbered according to Kabat et al., supra.

[0186] The phrases "Kabat variable domain residue numbering" or "Kabat amino acid position numbering" and variations thereof refer to the numbering system used for the heavy chain variable domain or light chain variable domain of the antibody compilation in Kabat et al., supra. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortening of, or insertion into, the FRs or HVRs of the variable domain. For example, a heavy chain variable domain may contain a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and inserted residues after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat numbering of residues can be determined for a given antibody by aligning the antibody sequence at the regions of homology with the "standard" Kabat numbering sequence.

[0187] "Framework" or "FR" residues are those variable domain residues other than the HVR residues as herein defined.

[0188] A "human consensus framework" or "acceptor human framework" is a framework representing the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991). Specific examples include the following: for VL, the subgroup can be subgroup kappa I, kappa II, kappa III, or kappa IV as in Kabat et al., supra; and for VH, the subgroup can be subgroup I, subgroup II, or subgroup III as in Kabat et al., supra. Alternatively, a human consensus framework can be derived from the above specific residues, such as when human framework residues are selected based on their homology to the donor framework by aligning the donor framework sequence with a collection of different human framework sequences. An acceptor human framework "derived from" a human immunoglobulin framework or human consensus framework may comprise the same amino acid sequence or may contain pre-existing amino acid sequence changes. In some embodiments, the number of pre-existing amino acid changes is 10 or less, or 9 or less, or 8 or less, or 7 or less, or 6 or less, or 5 or less, or 4 or less, or 3 or less, or 2 or less.

[0189] A "VH subgroup III consensus framework" comprises a consensus sequence obtained from the amino acid sequences in variable heavy subgroup III of Kabat et al., supra. In one embodiment, the amino acid sequence of the VH subgroup III consensus framework comprises at least a portion or all of each of the following sequences: EVQLVESGGGLVQPGGSLRLSCAAS(HC-FR1) (SEQ ID NO: 25), WVRQAPGKGLEWV(HC-FR2) (SEQ ID NO: 26), RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (HC-FR3) (SEQ ID NO: 27), WGQGTLVTVSA(HC-FR4) (SEQ ID NO: 28).

[0190] A "VL kappa I consensus framework" comprises a consensus sequence obtained from the amino acid sequences of variable light kappa subgroup I of Kabat et al., supra. In one embodiment, the amino acid sequence of the VH subgroup I consensus framework comprises at least a portion or all of each of the following sequences: DIQMTQSPSSLSASVGDRVTITC(LC-FR1) (SEQ ID NO: 29), WYQQKPGKAPKLLIY(LC-FR2) (SEQ ID NO: 30), GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC(LC-FR3) (SEQ ID NO: 31), FGQGTKVEIKR(LC-FR4) (SEQ ID NO: 32).

[0191] For example, an "amino acid modification" at a particular position in the Fc region refers to the substitution or deletion of the particular residue, or the insertion of at least one amino acid residue adjacent to the particular residue. An insertion "adjacent to" a particular residue refers to an insertion within 1 to 2 residues thereof. The insertion may be N-terminal or C-terminal to the particular residue. The preferred amino acid modification here is substitution.

[0192] An "affinity matured" antibody is one with one or more alterations in one or more HVRs thereof that result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody that does not possess those alterations. In one embodiment, the affinity matured antibody has nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are produced by procedures known in the art. For example, Marks et al., Bio / Technology, 10:779-783 (1992) describe affinity maturation by shuffling VH and VL domains. Random mutagenesis of HVR and / or framework residues has been described, for example, by Barbas et al., Proc Nat Acad. Sci, USA 91:3809-3813 (1994); Schier et al., Gene, 169:147-155 (1995); Yelton et al., J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al., J. Mol. Biol. 226:889-896 (1992).

[0193] As used herein, the terms "specifically bind" or "specific for" refer to a measurable and reproducible interaction, such as binding between a target and an antibody, that determines the presence of the target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody that specifically binds to a target (which may be an epitope) is an antibody that binds to this target with greater affinity, avidity, ease, and / or duration than it binds to other targets. In one embodiment, the extent of binding of an antibody to an unrelated target is less than about 10% of the binding of the antibody to the target, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (Kd) of ≦1 μM, ≦100 nM, ≦10 nM, or ≦0.1 nM. In certain embodiments, an antibody specifically binds to an epitope on a protein that is conserved among proteins from different species. In another embodiment, specific binding can include, but does not require, exclusive binding.

[0194] As used herein, the term "immunoadhesin" refers to an antibody-like molecule that combines the binding specificity of a heterologous protein (an "adhesin") with the effector functions of immunoglobulin constant domains. Structurally, immunoadhesins comprise a fusion of an amino acid sequence with the desired binding specificity, which is other than the antigen recognition and binding site of an antibody (i.e., is "heterologous"), with an immunoglobulin constant domain sequence. The adhesin portion of the immunoadhesin molecule is typically a contiguous amino acid sequence that includes at least the binding site for a receptor or ligand. The immunoglobulin constant domain sequence of the immunoadhesin can be derived from any immunoglobulin, such as IgG-1, IgG-2 (including IgG2A and IgG2B), IgG-3, or IgG-4 subtypes, IgA (including IgA-1 and IgA-2), IgE, IgD, or IgM. Ig fusions preferably involve the substitution of an antibody or polypeptide domain as described herein in place of at least one variable region within an Ig molecule. In a particularly preferred embodiment, the immunoglobulin fusion comprises the hinge, CH2, and CH3, or the hinge, CH1, CH2, and CH3 regions of an IgG1 molecule. For information on the preparation of immunoglobulin fusions, see U.S. Patent No. 5,428,130, issued June 27, 1995. For example, immunoadhesins useful as second agents in the combination therapies described herein comprise a polypeptide comprising the extracellular or PD-1-binding portion of PD-L1 or PD-L2, or the extracellular or PD-L1 or PD-L2-binding portion of PD-1, fused to a constant domain of an immunoglobulin sequence, such as PD-L1 ECD-Fc, PD-L2 ECD-Fc, and PD-1 ECD-Fc, respectively. Immunoadhesin combinations of Ig Fc and ECD of cell surface receptors are sometimes referred to as soluble receptors.

[0195] "Fusion protein" and "fusion polypeptide" refer to a polypeptide having two portions covalently linked together, each of which is a polypeptide having different properties. The property can be a biological property, such as in vitro or in vivo activity. The property can also be simply a chemical or physical property, such as binding to a target molecule, catalysis of a reaction, etc. The two portions can be directly linked by a single peptide bond or through a peptide linker, but are in reading frame with each other.

[0196] A "PD-1 oligopeptide," "PD-L1 oligopeptide," or "PD-L2 oligopeptide" is an oligopeptide that binds, preferably specifically, to a PD-1, PD-L1, or PD-L2 negative costimulatory polypeptide, respectively, and includes a receptor, ligand, or signaling component described herein. Such oligopeptides can be chemically synthesized using known oligopeptide synthesis methods, or can be prepared and purified using recombinant techniques. Such oligopeptides typically are at least about 5 amino acids in length, or at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 , 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids in length or more. Such oligopeptides can be identified using well known techniques.In this regard, it should be noted that techniques for screening oligopeptide libraries for oligopeptides capable of specifically binding to a polypeptide target are well known in the art (e.g., U.S. Pat. Nos. 5,556,762, 5,750,373, 4,708,871, 4,833,092, 5,223,409, 5,403,484, 5,571,689, 5,663,143, PCT Publication Nos. WO 84 / 03506 and WO 84 / 03564; Geysen et al., Proc. Natl. Acad. Sci. USA, 81:3998-4002 (1984); Geysen et al., Proc. Natl. Acad. Sci. USA, 82:178-182 (1985); Geysen et al., Synthetic Peptides as Antigens, 130-149 (1989)). (1986); Geysen et al., J. Immunol. Meth., 102:259-274 (1987); Schoofs et al., J. Immunol., 140:611-616 (1988); Cwirla, S. E. et al., Proc. Natl. Acad. Sci. USA, 87:6378 (1990); Lowman, H. B. et al., Biochemistry, 30:10832 (1991); Clackson, T. et al., Nature, 352:624 (1991); Marks, J. D. et al., J. Mol. Biol., 222:581 (1991); Kang, A. S. et al., Proc. Natl. Acad. Sci. USA, 88:8363 (1991), and Smith, G. P., Current Opin. Biotechnol., 2:668 (1991)).

[0197] A "blocking" or "antagonist" antibody is one that inhibits or reduces the biological activity of the antigen to which it binds. In some embodiments, a blocking or antagonist antibody substantially or completely inhibits the biological activity of the antigen. The anti-PD-L1 antibodies of the invention block signaling through PD-1, thereby restoring a functional response by T cells (e.g., proliferation, cytokine production, target cell killing) to dysfunctional antigen stimulation.

[0198] An "agonist" or activating antibody is one that enhances or initiates signaling by the antigen to which it binds. In some embodiments, an agonist antibody causes or activates signaling in the absence of a natural ligand.

[0199] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy chain Fc region is usually defined to stretch from the amino acid residue at position Cys226 or Pro230 to the carboxyl terminus thereof. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) can be removed, for example, during antibody production or purification, or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Thus, an intact antibody composition can include antibodies with all K447 residues removed, antibodies with all K447 residues removed, and antibodies with a mixture of antibodies with and without the K447 residue. Suitable native-sequence Fc regions for use in the antibodies of the present invention include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4.

[0200] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. A preferred FcR is a native-sequence human FcR. Furthermore, a preferred FcR is one that binds an IgG antibody (a gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA (an "activating receptor") and FcγRIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains. Activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (see M. Daeron, Annu. Rev. Immunol., 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those identified in the future, are encompassed by the term "FcR" herein.

[0201] The term "Fc receptor" or "FcR" also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgG to the fetus. Guyer et al., J. Immunol. 117:587 (1976), and Kim et al., J. Immunol. 24:249 (1994). Methods for measuring binding to FcRn are known (see, e.g., Ghetie and Ward, Immunology Today, 18(12):592-8 (1997); Ghetie et al., Nature Biotechnology, 15(7):637-40 (1997); Hinton et al., J. Biol. Chem., 279(8):6213-6 (2004); WO 2004 / 92219 (Hinton et al.)). The in vivo binding to human FcRn and serum half-life of a human FcRn high-affinity-binding polypeptide can be assayed, for example, in transgenic mice or transfected human cell lines expressing human FcRn, or in primates administered with a polypeptide having a mutant Fc region. WO 2000 / 42072 (Presta) describes antibody variants with improved or decreased binding to FcRs. See also, for example, Shields et al., J. Biol. Chem., 9(2): 6591-6604 (2001).

[0202] As used herein, the phrases "substantially reduced" or "substantially different" refer to a sufficiently high degree of difference between two numerical values ​​(typically one associated with a molecule and the other associated with a reference / comparator molecule) that one of skill in the art would consider there to be a statistically significant difference between the two values ​​within the range of the biological characteristic measured by the values ​​(e.g., Kd values). The difference between the two values ​​may be, for example, about 10% or more, about 20% or more, about 30% or more, about 40% or more, and / or about 50% or more as a function of the value for the reference / comparator molecule.

[0203] As used herein, the terms "substantially similar" or "substantially the same" refer to a sufficiently high degree of similarity between two numerical values ​​(e.g., one associated with an antibody of the invention and the other associated with a reference / comparator antibody) that one of skill in the art would consider there to be little or no biologically and / or statistically significant difference between the two values ​​within the range of the biological characteristic measured by the value (e.g., Kd value). The difference between the two values ​​may be, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and / or less than about 10% as a function of the value for the reference / comparator molecule.

[0204] As used herein, "carrier" includes pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. Physiologically acceptable carriers are often aqueous pH-buffered solutions. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acid salts; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or nonionic surfactants, such as TWEEN. TM , polyethylene glycol (PEG), and PLURONICS TM Includes.

[0205] "Package insert" means instructions customarily included in commercial packaging of a pharmaceutical product, including information about the indications, uses, dosage, administration, contraindications, other medications that may be combined with the packaged product, and / or instructions customarily included in commercial packaging of a pharmaceutical product, including information about precautions regarding the use of such medications, etc.

[0206] As used herein, the term "treatment" refers to a clinical intervention designed to alter the natural course of the individual or cells being treated during the course of clinical pathology. Desirable effects of treatment include slowing the rate of disease progression, alleviating or ameliorating the disease state, and remission or improved prognosis. For example, an individual is successfully "treated" if one or more symptoms associated with cancer are alleviated or eliminated, including, but not limited to, reducing (or destroying) the proliferation of cancer cells, reducing symptoms caused by the disease, improving the quality of life of those affected by the disease, reducing the dose of other drugs required to treat the disease, slowing the progression of the disease, and / or prolonging the individual's survival.

[0207] As used herein, "delaying disease progression" means postponing, preventing, slowing, preventing, stabilizing, and / or postponing the onset of a disease (e.g., cancer). This delay can be for a variety of periods of time, depending on the disease being treated and / or the individual's medical history. As will be apparent to one of skill in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease. For example, late-stage cancer, such as the development of metastases, can be delayed.

[0208] As used herein, "reducing or inhibiting cancer recurrence" means reducing or inhibiting the recurrence of a tumor or cancer or the progression of a tumor or cancer.

[0209] As used herein, the terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Included in this definition are benign and malignant cancers, as well as dormant tumors or micrometastases. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include squamous cell carcinoma, lung cancer (including small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung), cancer of the peritoneum, hepatocellular carcinoma, gastric or abdominal cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, liver cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, and various types of head and neck cancer, and B-cell lymphoma (low-grade / follicular non-Hodgkin's lymphoma) lymphoma (NHL); small lymphocytic (SL) NHL; intermediate-grade / follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphoblastic NHL; high-grade small non-dividing cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); hairy cell leukemia; chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal blood vessel proliferation associated with phacomatosis, edema (e.g., associated with brain tumors), and Meigs' syndrome.

[0210] As used herein, "metastasis" refers to the spread of cancer from its primary site to other parts of the body. Cancer cells can break away from the primary tumor, infiltrate into lymphatics and blood vessels, circulate through the bloodstream, and grow (metastasize) at distant foci in normal tissues elsewhere in the body. Metastasis can be local or distant. Metastasis is a sequential process involving tumor cells breaking off from the primary tumor, traveling through the bloodstream, and terminating at a distant site. At the new site, the cells establish a blood supply and may grow to form a life-threatening mass. Both stimulatory and inhibitory molecular pathways within tumor cells regulate this behavior, and interactions between tumor cells and host cells at distant sites are also significant.

[0211] An "effective amount" is at least the minimum concentration required to produce a measurable improvement or prevention of a particular disease. The effective amount may vary depending on factors such as the disease state, age, sex, and weight of the patient, and the ability of the antibody to elicit a desired response in an individual. An effective amount is also one in which the therapeutically beneficial effects outweigh any toxic or adverse effects of the treatment. For prophylactic use, beneficial or desired results include those that eliminate or reduce the risk, reduce the severity, or delay the onset of disease, including biochemical, histological, and / or behavioral symptoms of the disease, complications present during disease development, and intermediate pathological phenotypes. For therapeutic use, beneficial or desired results include clinical results such as reducing one or more symptoms caused by the disease, improving the quality of life of those suffering from the disease, reducing the dose of other drugs required to treat the disease, enhancing the effectiveness of other medications through targeting, delaying disease progression, and / or prolonging survival. In the case of cancer or tumors, an effective amount of an agent may have the effect of reducing the number of cancer cells; reducing tumor size; inhibiting (i.e., slowing to some extent or preferably stopping) the infiltration of cancer cells into surrounding organs; inhibiting (i.e., slowing to some extent or preferably stopping) tumor metastasis; inhibiting (i.e., slowing to some extent or preferably stopping) tumor growth to some extent; and / or alleviating to some extent one or more symptoms associated with the disease. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of an agent, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly achieve prophylactic or therapeutic treatment. As understood in a clinical context, an effective amount of an agent, compound, or pharmaceutical composition may or may not be achieved in conjunction with another agent, compound, or pharmaceutical composition. Thus, an "effective amount" can be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in combination with one or more other agents, a desired result is likely to be or is achieved.

[0212] As used herein, "in combination with" refers to the administration of one treatment modality in addition to another treatment modality. Thus, "in combination with" refers to the administration of one treatment modality before, during, and after the administration of the other treatment modality to an individual.

[0213] As used herein, "subject" means a mammal, including, but not limited to, a human or non-human mammal, such as a cow, horse, dog, sheep, or cat. Preferably, the subject is a human. A patient is also a subject herein.

[0214] As used herein, "complete response" or "CR" means the disappearance of all target lesions; "partial response" or "PR" means at least a 30% reduction in the sum of the longest diameters (SLD) of the target lesions, referenced to the baseline SLD; "stable disease" or "SD" means neither sufficient shrinkage of target lesions to qualify for PR nor sufficient increase to qualify for PD, referenced to the smallest SLD since treatment began.

[0215] As used herein, "progressive disease" or "PD" means at least a 20% increase in the SLD of a target lesion, referring to the smallest SLD recorded since treatment initiation or in the presence of one or more new lesions.

[0216] As used herein, "progression-free survival" (PFS) means the length of time during and after treatment during which the disease being treated (e.g., cancer) does not worsen. Progression-free survival can include the length of time a patient is in complete or partial remission, as well as the length of time a patient is experiencing stable disease.

[0217] As used herein, "objective response rate" (ORR) means the sum of the complete response (CR) rate and the partial response (PR) rate.

[0218] As used herein, "overall survival rate" means the proportion of individuals in a population who are likely to be alive after a specified period of time.

[0219] A "chemotherapeutic agent" is a compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents, such as thiotepa and cyclophosphamide (CYTOXAN®); alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, mesuredopa, and uredopa; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; acetogenins, especially blatta camptothecin and blattacinone; delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; camptothecin (including synthetic analogs topotecan (HYCAMTIN®), CPT-11 (irinotecan, CAMPTOSAR®, acetylcamptothecin, scopolectin, and 9-aminocamptothecin); bryostatin; pemetrexed; kallistatin; CC-1065 (including its azotocin, carzela) carzelesin and bizelesin, including synthetic analogs; podocillotoxin; podophyllic acid; teniposide; cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatins; duocarmycins (including synthetic analogs, KW-2189 and CBI-TM1); eluterobin; pancratistatin; TLK-286; CDP323, an oral alpha-4 integrin inhibitor; sarcodictiin; spongiostatin spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard;nitrosureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gamma 11, calicheamicin omega 11 (e.g., Nicolaou et al., Angew Chem. Intl. Ed. Engl. 33:183-186 (1994); dynemicins, including dynemicin A; esperamicin; and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including ADRIAMYCIN®, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, doxorubicin HCl liposome injection (DOXIL®), and deoxydoxorubicin), epilepsy, erythromycin ... Bicine, esorubicin, idalarubicin, marcelomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, queramycin, lodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate, gemcitabine (GEMZAR®), tegafur (UFTORAL®), capecitabine (XELODA®), epothilones, and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine;Pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, and imatinib (2-phenylaminopyrimidine derivatives), and other c-kit inhibitors; antiadrenal drugs such as aminoglutethimide, mitotane, trilostane; folate replenishers such as furoic acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; eniluracil; amsacrine; bestravcil; bisantrene; edatracene acetate; defofamine; demecolcine; diazicon; eflornithine; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidynin; maytansinoids such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; fenameth; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; schizofiran; spirogermanium; tenuazonic acid; triazicon; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, veracrine A, roridin A, and anguidine); urethane; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); thiotepa; taxoids, such as paclitaxel (TAXOL®), albumin-engineered nanoparticle formulations of paclitaxel (ABRAXANE; TM), and docetaxel (TAXOTERE®); chlorambucil; 6-thioguanine, mercaptopurine, methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine (VELBANR®); platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine (ONCOVIN®); oxaliplatin; leucovorin; vinorelbine (NAVELBINE®); Novant thoron; edatrexate; daunomycin; aminopterin; ibandronate; the topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing; and CHOP, an abbreviation for the combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, and oxaliplatin (ELOXATIN) in combination with 5-FU and leucovorin. TM ) and combinations of two or more of the above, such as FOLFOX, which is an abbreviation for a treatment using FOLFOX.

[0220] Also included in this definition are antihormonal agents, often in the form of systemic or systemic treatments, that act to modulate, reduce, block, or suppress the effects of hormones that can promote cancer growth. They may also be hormones themselves. Specific examples include antiestrogens and selective estrogen receptor modulators (SERMs), such as tamoxifen (including NOLVADEX® tamoxifen), raloxifene (EVISTA®), droloxifene, 4-hydroxytamoxifen, trioxifene, ketoxifene, LY117018, onapristone, and toremifene (FARESTON®); antiprogesterones; estrogen receptor downregulators (ERDs); estrogen receptor antagonists, such as fulvestrant (FASLODEX®); and agents that function to suppress or shut down the ovaries, such as luteinizing hormone-releasing hormone (LHRH) agonists. antiandrogens, such as leuprolide acetate (LUPRON® and ELIGARD®), goserelin acetate, buserelin acetate, and tripterelin; other antiandrogens, such as flutamide, nilutamide, and bicalutamide; and aromatase inhibitors, which inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as 4(5)-imidazoles, aminoglutethimide, megestrol acetate (MEGASE®), exemestane (AROMASIN®), formestane, fadrozole, vorozole (RIVISOR®), letrozole (FEMARA®), and anastrozole (ARIMIDEX®).Also included in this definition of chemotherapeutic agent are bisphosphonates, such as clodronate (e.g., BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), pamidronate (AREDIA®), tiludronate (SKELID®), or risedronate (ACTONEL®); and troxacitabine (1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in adherent cell proliferation, such as PKC-alpha, Raf, H-Ras, and epidermal growth factor receptor (EGF-R); vaccines, such as THERATOPE® ) vaccines and gene therapy vaccines, such as ALLOVECTIN® vaccine, LEUVECTIN® vaccine, and VAXID® vaccine; topoisomerase 1 inhibitors (e.g., LURTOTECAN®); antiestrogens such as fulvestrant; Kit inhibitors such as imatinib or EXEL-0862 (tyrosine kinase inhibitors); EGFR inhibitors, such as erlotinib or cetuximab; anti-VEGF inhibitors, such as bevacizumab; alinotecan; rmRH (e.g., ABARELIX®); lapatinib and lapatinib ditosylate (an ErbB-2 and EGFR dual tyrosine kinase small molecule inhibitor, also known as GW572016); 17AAG (a geldanamycin derivative, which is a heat shock protein (HSP) 90 poison), and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing.

[0221] As used herein, the term "cytokine" generally refers to a protein released by one cell population that acts as an intercellular mediator on other cells or has an autocrine effect on the protein-producing cell. Examples of such cytokines include lymphokines, monokines; interleukins ("IL") such as IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-17A-F, IL-18 to IL-29 (e.g., IL-23), IL-31, including PROLEUKIN® rIL-2; tumor necrosis factors, e.g., TNF-α or TNF-β, TGF-β1-3; and other polypeptide factors, including leukemia inhibitory factor ("LIF"), ciliary neurotrophic factor ("CNTF"), CNTF-like cytokine ("CLC"), cardiotrophin ("CT"), and Kit Ligand ("KL").

[0222] As used herein, the term "chemokine" refers to soluble factors (e.g., cytokines) that have the ability to selectively induce chemotaxis and activation of leukocytes. They also initiate the processes of angiogenesis, inflammation, wound healing, and tumorigenesis. Examples of chemokines include IL-8, the human homolog of mouse keratinocyte-attractant (KC).

[0223] As used in the specification and the appended claims, the singular forms "a," "or," and "the" include plural referents unless the context clearly dictates otherwise.

[0224] By "about," a value or parameter herein includes (or describes) a variation that is directed to the value or parameter itself. For example, a statement referring to "about X" includes the statement of "X."

[0225] As used herein, the phrase "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of a compound of the present invention. Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate "mesylate", ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts, alkali metal (e.g., sodium and potassium) salts, alkaline earth metal (e.g., magnesium) salts, and ammonium salts. A pharmaceutically acceptable salt may involve the inclusion of other molecules such as acetate ions, succinate ions, or other counter ions. A counter ion may be any organic or inorganic moiety that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. When multiple charged atoms are part of the pharmaceutically acceptable salt, it may have multiple counter ions. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counter ions.

[0226] When the compound of the present invention is a base, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, and the like, or with an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, e.g., glucuronic acid or galacturonic acid, an alpha hydroxy acid, e.g., citric acid or tartaric acid, an amino acid, e.g., aspartic acid or glutamic acid, an aromatic acid, e.g., benzoic acid or cinnamic acid, a sulfonic acid, e.g., p-toluenesulfonic acid or ethanesulfonic acid, and the like.

[0227] If the compound of the invention is an acid, the desired pharmaceutically acceptable salt can be prepared by any suitable method, such as treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary), an alkali metal hydroxide, or an alkaline earth metal hydroxide. Illustrative examples of suitable salts include, but are not limited to, organic bases derived from amino acids such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines such as piperidine, morpholine, and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

[0228] The term "pharmaceutically acceptable" indicates that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal being treated therewith.

[0229] It is understood that the aspects and variations of the invention described herein include those "consisting of" and / or "consisting essentially of" aspects and variations.

[0230] III. Method In one aspect, provided herein is a method for treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity.

[0231] In another aspect, provided herein is a method for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. As disclosed herein, cancer recurrence and / or cancer progression includes, but is not limited to, cancer metastasis.

[0232] In another aspect, provided herein is a method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that decreases or inhibits TIGIT expression and / or activity.

[0233] In another aspect, provided herein is a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity.

[0234] In some embodiments, the immune-related disease is associated with a T cell dysfunction disorder. In some embodiments, the immune-related disease is a viral infection. In certain embodiments, the viral infection is a chronic viral infection. In some embodiments, the T cell dysfunction disorder is characterized by a decreased response to antigenic stimulation. In some embodiments, the T cell dysfunction disorder is characterized by T cell anergy or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the T cells are CD4+ and CD8+ T cells. In some embodiments, the T cell dysfunction disorder includes unresolved acute infection, chronic infection, and tumor immunity.

[0235] In another aspect, provided herein is a method of increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to an individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that decreases or inhibits TIGIT expression and / or activity.

[0236] In another aspect, provided herein is a method for treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0237] In another aspect, provided herein is a method of reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0238] In another aspect, provided herein is a method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0239] In another aspect, provided herein is a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0240] In some embodiments, the immune-related disease is associated with a T cell dysfunction disorder. In some embodiments, the immune-related disease is a viral infection. In certain embodiments, the viral infection is a chronic viral infection. In some embodiments, the T cell dysfunction disorder is characterized by a decreased response to antigenic stimulation. In some embodiments, the T cell dysfunction disorder is characterized by T cell anergy, or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the T cells are CD4+ and CD8+ T cells. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

[0241] In another aspect, provided herein is a method of increasing, enhancing, or stimulating an immune response or function in an individual by administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0242] In some embodiments, an agent that modulates CD226 expression and / or activity can increase and / or stimulate CD226 expression and / or activity; increase and / or stimulate the interaction of CD226 with PVR, PVRL2, and / or PVRL3; and increase and / or stimulate intracellular signaling mediated by CD226 binding to PVR, PVRL2, and / or PVRL3. As used herein, an agent that can increase and / or stimulate CD226 expression and / or activity includes, but is not limited to, an agent that increases and / or stimulates CD226 expression and / or activity. As used herein, an agent that can increase and / or stimulate the interaction of CD226 with PVR, PVRL2, and / or PVRL3 includes, but is not limited to, an agent that increases and / or stimulates the interaction of CD226 with PVR, PVRL2, and / or PVRL3. As used herein, an agent that can increase and / or stimulate intracellular signaling mediated by the binding of CD226 to PVR, PVRL2, and / or PVRL3 includes, but is not limited to, an agent that increases and / or stimulates intracellular signaling mediated by the binding of CD226 to PVR, PVRL2, and / or PVRL3.

[0243] In some embodiments, the agent that modulates CD226 expression and / or activity is selected from an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof.

[0244] In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or antigen-binding fragment thereof. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an inhibitory nucleic acid selected from an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera.

[0245] In some embodiments, the antagonist of TIGIT expression and / or activity is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide. In some embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or antigen-binding fragment thereof. In some embodiments, the antagonist of TIGIT expression and / or activity is an inhibitory nucleic acid selected from an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera.

[0246] In some embodiments, the antagonist of PVR expression and / or activity is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In some embodiments, the antagonist of PVR expression and / or activity is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0247] In some embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVR is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In some embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVR is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0248] In some embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0249] In some embodiments, the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0250] In some embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In some embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0251] In some embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2 is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0252] In some embodiments, the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3 is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

[0253] In another aspect, provided herein are methods of increasing, enhancing, or stimulating an immune response or function in an individual by administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits expression and / or activity of one or more additional immune co-inhibitory receptors. In some embodiments, the one or more additional immune co-inhibitory receptors are selected from PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, and CD96. In some embodiments, the one or more additional immune co-inhibitory receptors are selected from PD-1, CTLA-4, LAG3, and TIM3.

[0254] In another aspect, provided herein are methods for increasing, enhancing, or stimulating an immune response or function in an individual by administering to the individual an effective amount of an agent that decreases or inhibits TIGIT expression and / or activity and an agent that increases or activates the expression and / or activity of one or more additional immune costimulatory receptors. In some embodiments, the one or more additional immune costimulatory receptors are selected from CD226, OX-40, CD28, CD27, CD137, HVEM, GITR, MICA, ICOS, NKG2D, and 2B4. In some embodiments, the one or more additional immune costimulatory receptors are selected from CD226, OX-40, CD28, CD27, CD137, HVEM, and GITR. In some embodiments, the one or more additional immune costimulatory receptors are selected from OX-40 and CD27.

[0255] The methods of the invention may find use in the treatment of conditions where enhanced immunogenicity is desired, such as increasing tumor immunogenicity for the treatment of cancer or T-cell dysfunction disorders.

[0256] A variety of cancers can be treated or their progression slowed.

[0257] In some embodiments, the individual has non-small cell lung cancer. The non-small cell lung cancer can be early or late stage. In some embodiments, the individual has small cell lung cancer. The small cell lung cancer can be early or late stage. In some embodiments, the individual has renal cell carcinoma. The renal cell carcinoma can be early or late stage. In some embodiments, the individual has colorectal cancer. The colorectal cancer can be early or late stage. In some embodiments, the individual has ovarian cancer. The ovarian cancer can be early or late stage. In some embodiments, the individual has breast cancer. The breast cancer can be early or late stage. In some embodiments, the individual has pancreatic cancer. The pancreatic cancer can be early or late stage. In some embodiments, the individual has gastric cancer. The gastric cancer can be early or late stage. In some embodiments, the individual has bladder cancer. The bladder cancer can be early or late stage. In some embodiments, the individual has esophageal cancer. The esophageal cancer can be early or late stage. In some embodiments, the individual has mesothelioma. The mesothelioma can be early or late stage. In some embodiments, the individual has melanoma. The melanoma can be early or late stage. In some embodiments, the individual has head and neck cancer. The head and neck cancer can be early or late stage. In some embodiments, the individual has thyroid cancer. The thyroid cancer can be early or late stage. In some embodiments, the individual has sarcoma. The sarcoma can be early or late stage. In some embodiments, the individual has prostate cancer. The prostate cancer can be early or late stage. In some embodiments, the individual has glioblastoma. The glioblastoma can be early or late stage. In some embodiments, the individual has cervical cancer. The cervical cancer can be early or late stage. In some embodiments, the individual is afflicted with thymic carcinoma. The thymic carcinoma can be at early or late stage. In some embodiments, the individual is afflicted with leukemia. The leukemia can be at early or late stage. In some embodiments, the individual is afflicted with lymphoma. The lymphoma can be at early or late stage. In some embodiments, the individual is afflicted with myeloma. The myeloma can be at early or late stage. In some embodiments, the individual is afflicted with mycosis fungoides.The mycosis fungoides can be at an early stage or a late stage. In some embodiments, the individual is afflicted with Merkel cell carcinoma. The Merkel cell carcinoma can be at an early stage or a late stage. In some embodiments, the individual is afflicted with a hematological malignancy. The hematological malignancy can be at an early stage or a late stage. In some embodiments, the individual is human.

[0258] In some embodiments of the methods of the invention, CD4 and / or CD8 T cells in the individual have increased or enhanced priming, activation, proliferation, cytokine release and / or cytolytic activity relative to before administration of the combination.

[0259] In some embodiments of the methods of the invention, the number of CD4 and / or CD8 T cells is increased relative to before administration of the combination. In some embodiments of the methods of the invention, the number of activated CD4 and / or CD8 T cells is increased relative to before administration of the combination.

[0260] In some embodiments of the methods of the invention, the activated CD4 and / or CD8 T cells are activated by γ-IFN. + The combination is characterized by increased CD4 production and / or increased cytolytic activity relative to pre-administration.

[0261] In some embodiments of the methods of the invention, the CD4 and / or CD8 T cells exhibit increased release of a cytokine selected from the group consisting of IFN-γ, TNF-α, and an interleukin.

[0262] In some embodiments of the methods of the invention, the CD4 and / or CD8 T cells are effector memory T cells. In some embodiments of the methods of the invention, the CD4 and / or CD8 effector memory T cells are γ-IFN-. + In some embodiments of the methods of the invention, the CD4 and / or CD8 effector memory T cells are characterized by CD44 and / or CD8 T cells and / or enhanced cytolytic activity. high CD62L low The gene is characterized by exhibiting expression of

[0263] In some embodiments of the methods of the invention, the cancer has an increased level of T cell infiltration.

[0264] In some embodiments, the methods of the present invention may further comprise administering an additional therapy. The additional therapy may be radiation therapy, surgery, chemotherapy, gene therapy, DNA therapy, viral therapy, RNA therapy, immunotherapy, bone marrow transplant, nanotherapy, monoclonal antibody therapy, or a combination thereof. The additional therapy may be in the form of adjuvant or neoadjuvant therapy. In some embodiments, the additional therapy is the administration of a side effect limiting agent (e.g., an agent intended to reduce the occurrence and / or severity of side effects of the treatment, such as an antiemetic). In some embodiments, the additional therapy is radiation therapy. In some embodiments, the additional therapy is surgery. In some embodiments, the additional therapy may be one or more of the chemotherapeutic agents described above.

[0265] Any of the PD-1 axis binding antagonists and agents that decrease or inhibit TIGIT expression and / or activity described below can be used in the methods of the invention.

[0266] In some embodiments, any of the targets described herein (e.g., PD-1, PD-L1, PD-L2, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, CD96, B7-1, TIGIT, CD226, OX-40, CD28, CD27, CD137, HVEM, GITR, MICA, ICOS, NKG2D, 2B4, etc.) is a human protein.

[0267] PD-1 axis binding antagonists Provided herein are methods for treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein are methods for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein are methods for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein is a method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein is a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis-binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity.

[0268] For example, PD-1 axis binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0269] In some embodiments, the PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its ligand binding partner. In particular aspects, the PD-1 ligand binding partner is PD-L1 and / or PD-L2. In other embodiments, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In particular aspects, the PD-L1 binding partner is PD-1 and / or B7-1. In other embodiments, the PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to its binding partner. In particular aspects, the PD-L2 binding partner is PD-1. The antagonist can be an antibody, antigen-binding fragment thereof, immunoadhesin, fusion protein, or oligopeptide.

[0270] In some embodiments, the PD-1 binding antagonist is selected from MDX-1106 (nivolumab), Merck 3745 (lambrolizumab), CT-011 (pidilizumab), and AMP-224. In some embodiments, the PD-L1 binding antagonist is selected from YW243.55.S70, MPDL3280A, MDX-1105, and MEDI4736. In some embodiments, the PD-L2 binding antagonist is AMP-224. In some embodiments, the PD-1 binding antagonist is AMP-224. MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in WO 2007 / 005874. Antibody YW243.55.S70 (SEQ ID NO: 20) is an anti-PD-L1 antibody described in WO 2010 / 077634A1 and U.S. Patent No. 8,217,149, which are incorporated herein by reference. MDX-1106, also known as MDX-1106-04, ONO-4538, BMS-936558, or nivolumab, is an anti-PD-1 antibody described in WO 2006 / 121168. Merck 3745, also known as MK3475, MK-3475, SCH-900475, or lambrolizumab, is an anti-PD-1 antibody described in WO 2009 / 114335. CT-011, also known as hBAT, hBAT-1, or pidilizumab, is an anti-PD-1 antibody described in WO 2009 / 101611. AMP-224, also known as B7-DCIg, is a PD-L2-Fc fusion soluble receptor described in WO 2010 / 027827 and WO 2011 / 066342.

[0271] Examples of anti-PD-L1 antibodies useful in the methods of the invention, and methods for making them, are described in PCT Patent Application Publication No. WO 2010 / 077634 A1 and U.S. Patent No. 8,217,149, which are incorporated herein by reference.

[0272] In some embodiments, the PD-1 axis binding antagonist is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody can inhibit the binding between PD-L1 and PD-1 and / or between PD-L1 and B7-1. In some embodiments, the anti-PD-L1 antibody is a monoclonal antibody. In some embodiments, the anti-PD-L1 antibody is an antibody fragment selected from the group consisting of Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragments. In some embodiments, the anti-PD-L1 antibody is a humanized antibody. In some embodiments, the anti-PD-L1 antibody is a human antibody.

[0273] The anti-PD-L1 antibodies useful in the invention, including compositions comprising such antibodies, such as those described in WO 2010 / 077634 A1 and U.S. Patent No. 8,217,149, can be used in combination with agents that decrease or inhibit TIGIT expression and / or activity, with or without any additional therapy (e.g., chemotherapy), to treat cancer or immune-related diseases (e.g., T-cell dysfunction diseases, viral infections, chronic viral infections, etc.).

[0274] In one embodiment, the anti-PD-L1 antibody comprises a heavy chain variable region polypeptide comprising HVR-H1, HVR-H2, and HVR-H3 sequences, wherein: (a) the HVR-H1 sequence is GFTFSX1SWIH (SEQ ID NO: 33); (b) the HVR-H2 sequence is AWIX2PYGGSX3YYADSVKG (SEQ ID NO: 34); (c) the HVR-H3 sequence is RHWPGGFDY (SEQ ID NO: 19); Further, where X1 is D or G; X2 is S or L; and X3 is T or S.

[0275] In one particular embodiment, X1 is D; X2 is S, and X3 is T. In other embodiments, the polypeptide further comprises variable region heavy chain framework sequences juxtaposed between the HVRs according to the following formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4). In yet another embodiment, the framework sequences are derived from a human consensus framework sequence. In a further embodiment, the framework sequence is a VH subgroup III consensus framework. In yet a further embodiment, at least one of the framework sequences is: HC-FR1 is EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25), HC-FR2 is WVRQAPGKGLEWV (SEQ ID NO: 26), HC-FR3 is RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27); HC-FR4 is WGQGTLVTVSA (SEQ ID NO: 28).

[0276] In yet a further aspect, the heavy chain polypeptide is further combined with a variable region light chain comprising HVR-L1, HVR-L2, and HVR-L3, wherein: (a) the HVR-L1 sequence is RASQX4X5X6TX7X8A (SEQ ID NO: 35); (b) HVR-L2 sequence is SASX9LX 10 S (SEQ ID NO: 36); (c) HVR-L3 sequence is QQX 11 X 12 X 13 X 14 PX 15 T (SEQ ID NO: 37); Further, wherein X4 is D or V; X5 is V or I; X6 is S or N; X7 is A or F; X8 is V or L; X9 is F or T; 10 is Y or A; X 11 is Y, G, F, or S; X 12 is L, Y, F or W; X 13is Y, N, A, T, G, F, or I; X 14 is H, V, P, T or I; X 15 is A, W, R, P or T.

[0277] In still further embodiments, X4 is D; X5 is V; X6 is S; X7 is A; X8 is V; X9 is F; X10 is Y; X11 is Y; X12 is L; X13 is Y; X14 is H; and X15 is A. In still further embodiments, the light chain further comprises variable region light chain framework sequences juxtaposed between the HVRs according to the formula (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In still further embodiments, the framework sequences are derived from human consensus framework sequences. In still further embodiments, the framework sequences are VL kappa I consensus frameworks. In still further embodiments, at least one of the framework sequences is as follows: LC-FR1 is DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29), LC-FR2 is WYQQKPGKAPKLLIY (SEQ ID NO: 30), LC-FR3 is GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31), LC-FR4 is FGQGTKVEIKR (SEQ ID NO: 32).

[0278] In another embodiment, an isolated anti-PD-L1 antibody or antigen-binding fragment is provided, comprising a heavy chain and a light chain variable region sequence, wherein: (a) the heavy chain comprises HVR-H1, HVR-H2, and HVR-H3, and further comprises: (i) the HVR-H1 sequence is GFTFSX1SWIH (SEQ ID NO: 33) (ii) the HVR-H2 sequence is AWIX2PYGGSX3YYADSVKG (SEQ ID NO: 34) (iii) the HVR-H3 sequence is RHWPGGFDY (SEQ ID NO: 19) (b) the light chain comprises HVR-L1, HVR-L2, and HVR-L3, and further comprises: (i) the HVR-L1 sequence is RASQX4X5X6TX7X8A (SEQ ID NO: 35) (ii) HVR-L2 sequence is SASX9LX 10 S (SEQ ID NO: 36) (iii) HVR-L3 sequence is QQX 11 X 12 X 13 X 14 PX 15 T (SEQ ID NO: 37) Further wherein: X1 is D or G; X2 is S or L; X3 is T or S; X4 is D or V; X5 is V or I; X6 is S or N; X7 is A or F; X8 is V or L; X9 is F or T; 10 is Y or A; X 11 is Y, G, F or S; X 12 is L, Y, F or W; X 13 is Y, N, A, T, G, F or I; X 14 is H, V, P, T or I; X 15 is A, W, R, P or T.

[0279] In certain embodiments, X1 is D; X2 is S, and X3 is T. In other embodiments, X4 is D; X5 is V; X6 is S; X7 is A; X8 is V; X9 is F; and X 10 is Y;X 11 is Y;X 12 is L;X 13 is Y;X 14 is H;X 15 is A. In yet another embodiment, X1 is D; X2 is S and X3 is T, X4 is D; X5 is V; X6 is S; X7 is A; X8 is V; X9 is F; and X 10 is Y;X 11 is Y;X 12 is L;X 13 is Y;X 14 is H and X 15is A.

[0280] In a further aspect, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet a further aspect, the framework sequences are derived from a human consensus framework sequence. In yet a further aspect, the heavy chain framework sequence is derived from a Kabat subgroup I, II, or III sequence. In yet a further aspect, the heavy chain framework sequence is a VH subgroup III consensus framework. In yet a further aspect, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 26) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSA (sequence number: 28).

[0281] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIKR (sequence number: 32).

[0282] In yet another particular aspect, the antibody further comprises a human or murine constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the murine constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the murine constant region is IgG2A. In a still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function is due to an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0283] In yet another embodiment, an anti-PD-L1 antibody is provided comprising a heavy chain and light chain variable region sequence, wherein: (a) the heavy chain further comprises HVR-H1, HVR-H2, and HVR-H3 sequences having at least 85% sequence identity to GFTFSDSWIH (SEQ ID NO: 17), AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and RHWPGGFDY (SEQ ID NO: 19), respectively; or (b) the light chain further comprises HVR-L1, HVR-L2, and HVR-L3 sequences having at least 85% sequence identity to RASQDVSTAVA (SEQ ID NO: 20), SASFLYS (SEQ ID NO: 21), and QQYLYHPAT (SEQ ID NO: 22), respectively. (c) In certain embodiments, the sequence identity is 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In other embodiments, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet other embodiments, the framework sequences are derived from human consensus framework sequences. In yet a further embodiment, the heavy chain framework sequences are derived from Kabat subgroup I, II, or III sequences. In yet a further embodiment, the heavy chain framework sequences are VH subgroup III consensus frameworks. In yet a further embodiment, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 26) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSA (sequence number: 28).

[0284] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIKR (sequence number: 32).

[0285] In yet another particular aspect, the antibody further comprises a human or murine constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the murine constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the murine constant region is IgG2A. In a still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function is due to an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0286] In another further embodiment, there is provided an isolated anti-PD-L1 antibody comprising a heavy chain and a light chain variable region sequence, wherein: (a) The heavy chain sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23), EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 40), or EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 41) or has at least 85% sequence identity to (b) the light chain sequence: It has at least 85% sequence identity to DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24).

[0287] In particular aspects, the sequence identity is 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In other aspects, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet other aspects, the framework sequences are derived from human consensus framework sequences. In a further embodiment, the heavy chain framework sequences are derived from Kabat subgroup I, II, or III sequences. In yet a further embodiment, the heavy chain framework sequences are VH subgroup III consensus frameworks. In yet a further embodiment, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 26) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSA (sequence number: 28).

[0288] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIKR (sequence number: 32).

[0289] In yet another particular aspect, the antibody further comprises a human or murine constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the murine constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the murine constant region is IgG2A. In a still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function results from production in a prokaryotic cell. In a still further particular aspect, the minimal effector function results from an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0290] In another further embodiment, the invention provides a composition comprising any of the above-mentioned anti-PD-L1 antibodies in combination with at least one pharmaceutically acceptable carrier.

[0291] In another further embodiment, an isolated nucleic acid encoding an anti-PD-L1 antibody light chain or heavy chain variable region sequence is provided, wherein: (a) the heavy chain further comprises HVR-H1, HVR-H2, and HVR-H3 sequences having at least 85% sequence identity to GFTFSDSWIH (SEQ ID NO: 17), AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and RHWPGGFDY (SEQ ID NO: 19), respectively; and (b) the light chain further comprises HVR-L1, HVR-L2, and HVR-L3 sequences having at least 85% sequence identity to RASQDVSTAVA (SEQ ID NO: 20), SASFLYS (SEQ ID NO: 21), and QQYLYHPAT (SEQ ID NO: 22), respectively.

[0292] In certain embodiments, the sequence identity is 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet other embodiments, the framework sequences are derived from human consensus framework sequences. In a further embodiment, the heavy chain framework sequences are derived from Kabat subgroup I, II, or III sequences. In yet a further embodiment, the heavy chain framework sequences are VH subgroup III consensus frameworks. In yet a further embodiment, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 26) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSA (sequence number: 28).

[0293] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIKR (sequence number: 32).

[0294] In yet another particular aspect, the antibody further comprises a human or murine constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the murine constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the murine constant region is IgG2A. In another still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function results from production in a prokaryotic cell. In a still further particular aspect, the minimal effector function results from an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0295] In another further embodiment, provided is an isolated anti-PDL1 antibody comprising a heavy chain and a light chain variable region sequence, wherein: (a) The heavy chain sequence: has at least 85% sequence identity to EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO:41); or (b) the light chain sequence: It has at least 85% sequence identity to DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24).

[0296] In particular aspects, the sequence identity is 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In other aspects, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet other aspects, the framework sequences are derived from human consensus framework sequences. In a further embodiment, the heavy chain framework sequences are derived from Kabat subgroup I, II, or III sequences. In yet a further embodiment, the heavy chain framework sequences are VH subgroup III consensus frameworks. In yet a further embodiment, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 26) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSS (sequence number: 42).

[0297] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIKR (sequence number: 32).

[0298] In yet another particular aspect, the antibody further comprises a human or murine constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the murine constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the murine constant region is IgG2A. In a still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function results from production in a prokaryotic cell. In a still further particular aspect, the minimal effector function results from an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0299] In a further embodiment, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet a further embodiment, the framework sequences are derived from a human consensus framework sequence. In yet a further embodiment, the heavy chain framework sequence is derived from a Kabat subgroup I, II, or III sequence. In yet a further embodiment, the heavy chain framework sequence is a VH subgroup III consensus framework. In yet a further embodiment, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAASGFTFS (SEQ ID NO: 43) HC-FR2 WVRQAPGKGLEWVA (SEQ ID NO: 44) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSS (sequence number: 45).

[0300] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIK (sequence number: 46).

[0301] In yet another particular aspect, the antibody further comprises a human or mouse constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the mouse constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the mouse constant region is IgG2A. In a still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function is due to an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0302] In yet another embodiment, provided is an anti-PDL1 antibody comprising a heavy chain and a light chain variable region sequence, wherein: (d) the heavy chain further comprises HVR-H1, HVR-H2, and HVR-H3 sequences having at least 85% sequence identity to GFTFSDSWIH (SEQ ID NO: 17), AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and RHWPGGFDY (SEQ ID NO: 19), respectively; or (e) the light chain further comprises HVR-L1, HVR-L2, and HVR-L3 sequences having at least 85% sequence identity to RASQDVSTAVA (SEQ ID NO: 20), SASFLYS (SEQ ID NO: 21), and QQYLYHPAT (SEQ ID NO: 22), respectively.

[0303] In certain embodiments, the sequence identity is 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the heavy chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and the light chain variable region comprises one or more framework sequences juxtaposed between the HVRs, such as (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In yet other embodiments, the framework sequences are derived from human consensus framework sequences. In yet a further embodiment, the heavy chain framework sequences are derived from Kabat subgroup I, II, or III sequences. In yet a further embodiment, the heavy chain framework sequences are VH subgroup III consensus frameworks. In yet a further embodiment, the one or more heavy chain framework sequences are as follows: HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 25) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 26) HC-FR3 RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 27) HC-FR4 WGQGTLVTVSSASTK (sequence number: 47).

[0304] In yet a further embodiment, the light chain framework sequences are derived from a Kabat kappa I, II, II, or IV subgroup sequence. In yet a further embodiment, the light chain framework sequences are VL kappa I consensus frameworks. In yet a further embodiment, the one or more light chain framework sequences are as follows: LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 29) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 30) LC-FR3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 31) LC-FR4 FGQGTKVEIKR (sequence number: 32).

[0305] In yet another particular aspect, the antibody further comprises a human or mouse constant region. In a still further aspect, the human constant region is selected from the group consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further particular aspect, the human constant region is IgG1. In a still further aspect, the mouse constant region is selected from the group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still further aspect, the mouse constant region is IgG2A. In a still further particular aspect, the antibody has reduced or minimal effector function. In a still further particular aspect, the minimal effector function is due to an "effectorless Fc mutation" or aglycosylation. In another further embodiment, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region.

[0306] In another further embodiment, provided is an isolated anti-PDL1 antibody comprising a heavy chain and a light chain variable region sequence, wherein: (a) The heavy chain sequence: has at least 85% sequence identity to EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 40); or (b) the light chain sequence: It has at least 85% sequence identity to DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24).

[0307] In some embodiments, provided is an isolated anti-PDL1 antibody comprising a heavy chain and a light chain variable region sequence, wherein the light chain variable region sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:24. In some embodiments, provided is an isolated anti-PDL1 antibody comprising heavy and light chain variable region sequences, wherein the heavy chain variable region sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:40. In some embodiments, the antibody is an isolated anti-PDL1 antibody comprising heavy and light chain variable region sequences, wherein the light chain variable region sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:24, and the heavy chain variable region sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:40.

[0308] In another further embodiment, provided is an isolated anti-PDL1 antibody comprising a heavy chain and a light chain variable region sequence, wherein: (a) The heavy chain sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 48); or (b) the light chain sequence: It has at least 85% sequence identity to DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 49).

[0309] In some embodiments, provided is an isolated anti-PDL1 antibody comprising heavy and light chain sequences, wherein the light chain sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 49. In some embodiments, provided is an isolated anti-PDL1 antibody comprising heavy and light chain sequences, wherein the heavy chain sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 48. In some embodiments, provided is an isolated anti-PDL1 antibody comprising heavy and light chain sequences, wherein the light chain sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:49, and the heavy chain sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:48.

[0310] In still further embodiments, the nucleic acid further comprises a vector suitable for expressing the nucleic acid encoding any of the anti-PD-L1 antibodies described above. In yet another particular embodiment, the vector further comprises a host cell suitable for expressing the nucleic acid. In yet another particular embodiment, the host cell is a eukaryotic or prokaryotic cell. In yet another particular embodiment, the eukaryotic cell is a mammalian cell, such as a Chinese hamster ovary (CHO) cell.

[0311] Anti-PD-L1 antibodies or antigen-binding fragments thereof can be produced using methods known in the art, for example by methods comprising culturing a host cell containing nucleic acid encoding any of the above-described anti-PD-L1 antibodies or antigen-binding fragments, in a form suitable for expression, under conditions suitable to produce such antibodies or fragments, and recovering the antibodies or fragments.

[0312] In another further embodiment, the invention provides compositions comprising an anti-PD-L1 antibody, or antigen-binding fragment thereof, provided herein and at least one pharmaceutically acceptable carrier.

[0313] Agents that reduce or inhibit TIGIT expression and / or activity Provided herein is a method for treating or delaying the progression of cancer in an individual, the method comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein is a method for reducing or inhibiting cancer recurrence or cancer progression in an individual, the method comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein is a method for treating or delaying the progression of an immune-related disease in an individual, the method comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein is a method for reducing or inhibiting the progression of an immune-related disease in an individual, the method comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity. Also provided herein is a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist in combination with an agent that decreases or inhibits TIGIT expression and / or activity. Also provided herein is a method of increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional immune co-inhibitory receptors. Also provided herein is a method of increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional immune co-stimulatory receptors. For example, agents that reduce or inhibit TIGIT expression and / or activity include antagonists of TIGIT expression and / or activity, antagonists of PVR expression and / or activity, agents that inhibit and / or block the interaction of TIGIT with PVR, agents that inhibit and / or block the interaction of TIGIT with PVRL2, agents that inhibit and / or block the interaction of TIGIT with PVRL3, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVR, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL2, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof.

[0314] In some embodiments, antagonists of TIGIT expression and / or activity include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0315] In some embodiments, antagonists of PVR expression and / or activity include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0316] In some embodiments, agents that inhibit and / or block the interaction of TIGIT with PVR include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0317] In some embodiments, agents that inhibit and / or block the interaction of TIGIT with PVRL2 include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0318] In some embodiments, agents that inhibit and / or block the interaction of TIGIT with PVRL3 include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0319] In some embodiments, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVR include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0320] In some embodiments, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL2 include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0321] In some embodiments, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL3 include small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

[0322] In some embodiments, the antagonist of TIGIT expression and / or activity is an inhibitory nucleic acid selected from an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera.

[0323] In some embodiments, the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or antigen-binding fragment thereof.

[0324] Anti-TIGIT antibodies useful in the present invention may be used in combination with PD-1 axis binding antagonists, including compositions comprising such antibodies, such as those described in WO 2009 / 126688.

[0325] Anti-TIGIT antibody The present invention provides anti-TIGIT antibodies. Exemplary antibodies include polyclonal, monoclonal, humanized, bispecific, and heteroconjugate antibodies. The present invention also provides antibodies against other polypeptides (i.e., anti-PVR antibodies), and it will be understood by those skilled in the art that anything described herein that specifically relates to methods of making, producing, modifying, using, or other aspects of anti-TIGIT antibodies will also be applicable to antibodies specific for other non-TIGIT polypeptides.

[0326] Polyclonal antibodies The anti-TIGIT antibody may include a polyclonal antibody. Methods for preparing polyclonal antibodies are known to those skilled in the art. Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and / or adjuvant are injected into the mammal by multiple subcutaneous or intraperitoneal injections. The immunizing agent may comprise a TIGIT polypeptide or a fusion protein thereof. It may be useful to conjugate the immunizing agent to a protein known to be immunogenic in the mammal being immunized. Examples of such immunogen proteins include, but are not limited to, keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. Examples of adjuvants that may be used include Freund's complete adjuvant and MPL-TDM adjuvant (monophosphoryl lipid A, synthetic trehalose dicorynomycolate). The immunization protocol may be selected by one skilled in the art without undue experimentation.

[0327] Monoclonal antibodies Alternatively, the anti-TIGIT antibody may be a monoclonal antibody. Monoclonal antibodies can be prepared using hybridoma methods, such as those described in Kohler and Milstein, Nature, 256:495 (1975). In the hybridoma method, a mouse, hamster, or other suitable host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce, or are capable of producing, antibodies that specifically bind to the immunizing agent. Alternatively, lymphocytes can be immunized in vitro.

[0328] The immunizing agent typically contains the TIGIT polypeptide of interest or a fusion protein thereof. Generally, peripheral blood lymphocytes ("PBLs") are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell [Goding, Monoclonal Antibodies: Principles and Practice, Academic Press, (1986) pp. 59-103]. The immortalized cell line is usually a transformed mammalian cell, particularly a myeloma cell of rodent, bovine, or human origin. Rat or mouse myeloma cell lines are usually used. The hybridoma cells may be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the survival or growth of the unfused, immortalized cells. For example, if the parent cells lack the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT), the hybridoma culture medium typically contains hypoxanthine, aminopterin, and thymidine ("HAT medium"), which prevents the growth of HGPRT-deficient cells.

[0329] Preferred immortalized cell lines are those that fuse efficiently, support stable, high-level antibody expression by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are mouse myeloma lines, which are available, for example, from the Salk Institute Cell Distribution Center in San Diego, California, or the American Type Culture Collection in Manassas, Virginia. Human myeloma and mouse-human heteromyeloma cell lines for producing human monoclonal antibodies have also been described [Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., *Monoclonal Antibody Production Techniques and Applications*, Marcel Dekker, Inc., New York, (1987) pp. 51-63].

[0330] The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the polypeptide. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity of the monoclonal antibody can be determined, for example, by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0331] After the desired hybridoma cells are identified, the clones can be subcloned by limiting dilution procedures and grown by standard methods [Goding, supra]. Suitable media for this purpose include, for example, Dulbecco's modified Eagle's medium and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

[0332] The monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0333] Monoclonal antibodies can also be produced by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., using oligonucleotide probes capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA can be placed into an expression vector, which can then be transfected into host cells, such as monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not produce immunoglobulin proteins, to allow the synthesis of monoclonal antibodies in the recombinant host cells. The DNA can also be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains for the homologous murine sequences [U.S. Pat. No. 4,816,567; Morrison et al., supra], or by covalently linking all or part of the coding sequence for a non-immunoglobulin polypeptide to the immunoglobulin coding sequence. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-binding site of an antibody of the invention to create a chimeric bivalent antibody.

[0334] The antibody may be a monovalent antibody. Methods for preparing monovalent antibodies are well known in the art. For example, one method involves recombinant expression of an immunoglobulin light chain and a modified heavy chain. The heavy chain is generally truncated at a point in the Fc region to prevent cross-linking of the heavy chain. Alternatively, the relevant cysteine ​​residues are substituted with other amino acid residues or deleted to prevent cross-linking.

[0335] In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using conventional techniques known in the art.

[0336] Human and Humanized Antibodies Anti-TIGIT antibodies of the present invention may further include humanized or human antibodies. Humanized forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (e.g., Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibodies) in which residues from a complementarity-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies may also comprise residues found neither in the recipient antibody nor in the imported CDR or framework sequences. Generally, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or nearly all of the CDR regions correspond to those of a non-human immunoglobulin and all or nearly all of the FR regions are those of a human immunoglobulin consensus sequence. Optionally, the humanized antibody will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op Struct. Biol., 2:593-596 (1992)).

[0337] Methods for humanizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acid residues introduced into it from a non-human source. These non-human amino acid residues are often referred to as "import" residues, typically from an "import" variable domain. Humanization can essentially be performed by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody according to the method of Winter and coworkers [Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)]. Such "humanized" antibodies are thus chimeric antibodies (U.S. Pat. No. 4,816,567) in which substantially less than intact human variable domains have been substituted by the corresponding sequences from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.

[0338] Human antibodies can also be produced using various methods known in the art, including phage display libraries [Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991)]. The techniques of Cole et al. and Boerner et al. can also be used to prepare human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss. p. 77 (1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991)]. Similarly, human antibodies can be produced by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the following scientific publications: Marks et al., Bio / Technology 10, 779-783 (1992); Lonberg et al., Nature 368 856-859 (1994); Morrison, Nature 368, 812-13 (1994); Fishwild et al., Nature Biotechnology 14, 845-51 (1996); Neuberger, Nature Biotechnology 14, 826 (1996); Lonberg and Huszar, Intern. Rev. Immunol. 13 65-93 (1995).

[0339] Antibodies can also be affinity matured using known selection and / or mutagenesis techniques, such as those described above. Preferred affinity-matured antibodies have 5-fold, more preferably 10-fold, and even more preferably 20- or 30-fold higher affinity than the starting antibody (generally murine, humanized, or human) from which the mature antibody is prepared.

[0340] bispecific antibody Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens. In the present case, one of the binding specificities is for TIGIT and the other is for any other antigen, preferably a cell surface protein or receptor or receptor subunit.

[0341] Methods for producing bispecific antibodies are known in the art. Traditionally, recombinant production of bispecific antibodies is based on the coexpression of two immunoglobulin heavy / light chain pairs, where the two heavy chains have different specificities [Milstein and Cuello, Nature, 305:537-539 (1983)]. Due to the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) generate a potential mixture of 10 different antibody molecules, of which only one has the correct bispecific structure. Purification of the correct molecule is usually achieved by affinity chromatography steps. Similar procedures are disclosed in WO 93 / 08829, published May 13, 1993, and in Traunecker et al., EMBO J., 10:3655-3656 (1991).

[0342] Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. Fusions preferably are with immunoglobulin heavy-chain constant domains, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred that the first heavy-chain constant domain (CH1) containing the site necessary for light-chain binding be present in at least one of the fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. For further details on producing bispecific antibodies, see, e.g., Suresh et al., Methods in Enzymology, 121:210 (1986).

[0343] According to another method described in WO 96 / 27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers recovered from recombinant cell culture. The preferred interface comprises at least a part of the CH3 region of the antibody constant domains. In this method, one or more small amino acid side chains from the interface of a first antibody molecule are replaced with larger side chains (e.g., tyrosine or tryptophan). A complementary "cavity" of identical or similar size to the large side chain(s) is created at the interface of the second antibody molecule by replacing the large amino acid side chain with a smaller one (e.g., alanine or threonine). This provides a mechanism for increasing the yield of heterodimers over other unwanted end-products such as homodimers.

[0344] Bispecific antibodies can be prepared as full-length antibodies or antibody fragments (e.g., F(ab')2 bispecific antibodies). Techniques for producing bispecific antibodies from antibody fragments have also been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. Brennan et al., Science, 229:81 (1985) describe a procedure for proteolytic cleavage of intact antibodies to produce F(ab')2 fragments. These fragments are reduced in the presence of the dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The resulting Fab' fragments are then converted to thionitrobenzoate (TNB) derivatives. One of the Fab'-TNB derivatives is then reconverted to the Fab'-thiol by reduction with mercaptoethylamine and mixed with an equimolar amount of the other Fab'-TNB derivative to form the bispecific antibody. The resulting bispecific antibody can be used as an agent for the selective immobilization of enzymes.

[0345] Fab' fragments can be directly recovered from E. coli and chemically coupled to form bispecific antibodies. Shalaby et al., J. Exp. Med., 175:217-225 (1992) describe the production of fully humanized bispecific antibody F(ab')2 molecules. Each Fab' fragment is secreted separately from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody. The bispecific antibody thus formed can bind to cells overexpressing the ErbB2 receptor and normal human T cells, and can elicit the cytolytic activity of human cytotoxic lymphocytes against human breast tumor targets.

[0346] Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et al., J. Immunol. 148(5):1547-1553 (1992). The leucine zipper peptides from the Fos and Jun proteins were linked to the Fab' portions of two different antibodies by gene fusion. Antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be used for the production of antibody homodimers. The "diabody" technology described by Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993) provided another mechanism for making bispecific antibody fragments. The fragments comprise a light-chain variable domain (V) separated by a linker that is too short to allow pairing between the two domains on the same chain. L ) to the heavy chain variable domain (V H ) are bonded together. Therefore, the V H and V L The domain is complementary to the V of another fragment. L and V H The Fv domains are forced to pair and form two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (sFv) dimers has also been reported. See Gruber et al., J. Immunol. 152:5368 (1994).

[0347] Antibodies with more than two valencies are also contemplated. As a non-limiting example, trispecific antibodies can be prepared. See, e.g., Tutt et al., J. Immunol. 147:60 (1991).

[0348] Exemplary bispecific antibodies can bind to two different epitopes of a TIGIT polypeptide provided herein. Alternatively, an arm of the anti-TIGIT polypeptide can be linked to an arm that binds to a triggering molecule on leukocytes, such as a T cell receptor molecule (e.g., CD2, CD3, CD28, or B7), or an Fc receptor for IgG (FcγR), such as FcγRI (CD64), FcγRII (CD32), and FcγRIII (CD16), to focus cellular defense mechanisms on cells expressing a specific TIGIT polypeptide. Bispecific antibodies can also be used to localize cytotoxic agents to cells expressing a specific TIGIT polypeptide. These antibodies have a TIGIT-binding arm and an arm that binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Another bispecific antibody of interest binds a TIGIT polypeptide and also binds tissue factor (TF).

[0349] Heteroconjugate antibodies Heteroconjugate antibodies are also within the scope of the present invention. Heteroconjugate antibodies consist of two covalently linked antibodies. Such antibodies have been proposed, for example, to target immune system cells to unwanted cells [U.S. Pat. No. 4,676,980] and for the treatment of HIV infection [WO 91 / 00360; WO 92 / 200373; EP 03089]. It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate, and those disclosed, for example, in U.S. Pat. No. 4,676,980.

[0350] Effector function processing It may be desirable to modify the effector function of the antibody of the present invention to improve the antibody's effectiveness in, for example, cancer therapy. For example, cysteine ​​residues may be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated may have improved internalization capability and / or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., J. Exp. Med. 176: 1191-1195 (1992) and Shopes, J. Immunol. 148: 2918-2922 (1992). Homodimeric antibodies with improved anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al., Cancer research 53: 2560-2565 (1993). Alternatively, an antibody can be engineered to have two Fc regions, thereby improving complement lysis and ADCC capabilities. See Stevenson et al., Anti-Cancer Drug Design 3: 219-230 (1989).

[0351] In some embodiments, anti-TIGIT antibodies were generated that were hamster anti-mouse antibodies. Two antibodies, 10A7 and 1F4, also specifically bound to human TIGIT. The amino acid sequences of the light and heavy chains of the 10A7 antibody were determined using standard techniques. The light chain sequence of this antibody was: The heavy chain sequence of this antibody is DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13). EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15), where the complementarity determining regions (CDRs) of each chain are represented by bold text. Thus, CDR1 of the 10A7 light chain has the sequence KSSQSLYYSGVKENLLA (SEQ ID NO: 1), CDR2 of the 10A7 light chain has the sequence ASIRFT (SEQ ID NO: 2), and CDR3 of the 10A7 light chain has the sequence QQGINNPLT (SEQ ID NO: 3). CDR1 of the 10A7 heavy chain has the sequence GFTFSSFTMH (SEQ ID NO: 4), CDR2 of the 10A7 heavy chain has the sequence FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and CDR3 of the 10A7 heavy chain has the sequence RPLGHNTFDS (SEQ ID NO: 6).

[0352] The amino acid sequences of the light and heavy chains of the 1F4 antibody were also determined. The light chain sequence of this antibody is: DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14), and the heavy chain sequence of this antibody is EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16), where the complementarity determining regions (CDRs) of each chain are represented by bold text. Thus, CDR1 of the 1F4 light chain has the sequence RSSQSLVNSYGNTFLS (SEQ ID NO: 7), CDR2 of the 1F4 light chain has the sequence GISNRFS (SEQ ID NO: 8), and CDR3 of the 1F4 light chain has the sequence LQGTHQPPT (SEQ ID NO: 9). CDR1 of the 1F4 heavy chain has the sequence GYSFTGHLMN (SEQ ID NO: 10), CDR2 of the 1F4 heavy chain has the sequence LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and CDR3 of the 1F4 heavy chain has the sequence GLRGFYAMDY (SEQ ID NO: 12).

[0353] The nucleotide sequence encoding the 1F4 light chain is GATGTTGTGTTGACTCAAACTCCACTCTCCCTGTCTGTCAGCTTTGGAGATCAAGTTTCTATCTCTTGCAGGTCTAGTCAGAGTCTTGTAAACAGTTATGGGAACACCTTTTTGTCTTGGTACCTGCACAAGCCTGGCCAGTCTCCACAGCTCCTCATCTTTGGGATTTCCAACAGATTTTCTGGGGTGCCAGACAGGTTCAGTGGCAGTGGTTCAGGGACAGATTTCACACTCAAGATCAGCACAATAAAGCCTGAGGACTTGGGAATGTATTACTGCTTACAAGGTACGCATCAGCCTCCCACGTTCGGTCCTGGGACCAAGCTGGAGGTGAAA (SEQ ID NO: 38). GAGGTCCAGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGAACTTCAATGAAGATATCCTGCAAGGCTTCTGGTTACTCATTCACTGGCCATCTTATGAACTGGGTGAAGCAGAGCCATGGAAAGAACCTTGAGTGGATTGGACTTATTATTCCTTACAATGGTGGTACAAGCTATAACCAGAA It was determined to be GTTCAAGGGCAAGGCCACATTGACTGTAGACAAGTCATCCAGCACAGCCTACATGGAGCTCCTCAGTCTGACTTCTGATGACTCTGCAGTCTATTTCTGTTCAAGAGGCCTTAGGGGCTTCTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 39).

[0354] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR comprising an amino acid sequence selected from the amino acid sequences set forth in (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6), or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

[0355] In some embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody light chain comprises: DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13) or It contains the amino acid sequence set forth in DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14).

[0356] In some embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody heavy chain is EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15) or It contains the amino acid sequence set forth in EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16).

[0357] In some embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody light chain comprises: DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13) or DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14), and the antibody heavy chain EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15) or It contains the amino acid sequence set forth in EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16).

[0358] In some embodiments, in the anti-TIGIT antibody or antigen-binding fragment thereof, the antibody is selected from a humanized antibody, a chimeric antibody, a bispecific antibody, a heteroconjugate antibody, and an immunotoxin.

[0359] In some embodiments, the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR that is at least 90% identical to an HVR set forth in any of (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6), or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

[0360] In some embodiments, the anti-TIGIT antibody or fragment thereof is DIVMTQSPSSLAVSPGEKVTMTCKSSQSLYYSGVKENLLAWYQQKPGQSPKLLIYYASIRFTGVPDRFTGSGSGTDYTLTITSVQAEDMGQYFCQQGINNPLTFGDGTKLEIKR (SEQ ID NO: 13) or DVVLTQTPLSLSVSFGDQVSISCRSSQSLVNSYGNTFLSWYLHKPGQSPQLLIFGISNRFSGVPDRFSGSGSGTDFTLKISTIKPEDLGMYYCLQGTHQPPTFGPGTKLEVK (SEQ ID NO: 14), or EVQLVESGGGLTQPGKSLKLSCEASGFTFSSFTMHWVRQSPGKGLEWVAFIRSGSGIVFYADAVRGRFTISRDNAKNLLFLQMNDLKSEDTAMYYCARRPLGHNTFDSWGQGTLVTVSS (SEQ ID NO: 15) or It includes a light chain and / or a heavy chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in EVQLQQSGPELVKPGTSMKISCKASGYSFTGHLMNWVKQSHGKNLEWIGLIIPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLTSDDSAVYFCSRGLRGFYAMDYWGQGTSVTVSS (SEQ ID NO: 16).

[0361] Agents that modulate CD226 expression and / or activity Provided herein are methods for treating or delaying the progression of cancer in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. Also provided herein are methods for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. Also provided herein are methods for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. Also provided herein are methods for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity. Also provided herein is a method for increasing, enhancing, or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

[0362] For example, an agent that modulates CD226 expression and / or activity is an agent that can increase and / or stimulate CD226 expression and / or activity, increase and / or stimulate the interaction of CD226 with PVR, PVRL2, and / or PVRL3, and increase and / or stimulate intracellular signaling mediated by CD226 binding to PVR, PVRL2, and / or PVRL3. In some embodiments, an agent that can increase and / or stimulate CD226 expression and / or activity is an agent that increases and / or stimulates CD226 expression and / or activity. In some embodiments, an agent that can increase and / or stimulate the interaction of CD226 with PVR, PVRL2, and / or PVRL3 is an agent that increases and / or stimulates the interaction of CD226 with PVR, PVRL2, and / or PVRL3. In some embodiments, an agent that can increase and / or stimulate intracellular signaling mediated by the binding of CD226 to PVR, PVRL2, and / or PVRL3 is an agent that increases and / or stimulates intracellular signaling mediated by the binding of CD226 to PVR, PVRL2, and / or PVRL3.

[0363] In some embodiments, the agent that modulates CD226 expression and / or activity is selected from an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is selected from a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or antigen-binding fragment thereof. In some embodiments, the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an inhibitory nucleic acid selected from an antisense polynucleotide, an interfering RNA,...

Claims

1. A method for treating or delaying the progression of cancer in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that decreases or inhibits TIGIT expression and / or activity.

2. A method for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

3. A method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

4. A method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that reduces or inhibits TIGIT expression and / or activity.

5. 5. The method of claim 3 or 4, wherein the immune-related disease is associated with a T-cell dysfunction disease.

6. 6. The method of claim 5, wherein the T cell dysfunction disorder is characterized by a decreased response to antigenic stimulation.

7. 6. The method of claim 5, wherein the T cell dysfunction disorder is characterized by T cell anergy or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity.

8. 6. The method of claim 5, wherein the T cell dysfunction disease is characterized by T cell depletion.

9. 9. The method of claim 3, wherein the T cells are CD4+ and CD8+ T cells.

10. 10. The method of any one of claims 3 to 9, wherein the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

11. A method for increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that decreases or inhibits TIGIT expression and / or activity.

12. A method for treating or delaying the progression of cancer in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

13. A method for reducing or inhibiting cancer recurrence or cancer progression in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

14. A method for treating or delaying the progression of an immune-related disease in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

15. A method for reducing or inhibiting the progression of an immune-related disease in an individual, comprising administering to the individual effective amounts of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

16. 16. The method of claim 14 or 15, wherein the immune-related disease is associated with a T-cell dysfunction disease.

17. 17. The method of claim 16, wherein the T cell dysfunction disorder is characterized by a decreased response to antigenic stimulation.

18. 17. The method of claim 16, wherein the T cell dysfunction disorder is characterized by T cell anergy, or a decreased ability to secrete cytokines, proliferate, or perform cytolytic activity.

19. 17. The method of claim 16, wherein the T cell dysfunction disease is characterized by T cell depletion.

20. 20. The method of any one of claims 16 to 19, wherein the T cells are CD4+ and CD8+ T cells.

21. 21. The method of any one of claims 14 to 20, wherein the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

22. A method for increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of a PD-1 axis binding antagonist and an agent that modulates CD226 expression and / or activity.

23. 23. The method of any one of claims 12 to 22, wherein the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates CD226 expression and / or activity.

24. 24. The method of any one of claims 12 to 23, wherein the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates the interaction of CD226 with PVR.

25. The method of any one of claims 12 to 24, wherein the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates intracellular signaling mediated by binding of CD226 to PVR.

26. The method of any one of claims 12 to 25, wherein the agent that modulates CD226 expression and / or activity is selected from the group consisting of an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, an agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof.

27. The method of claim 26, wherein the agent that modulates CD226 expression and / or activity is an agent that inhibits and / or blocks the interaction of CD226 with TIGIT.

28. The method of claim 26 or 27, wherein the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide.

29. The method of claim 26 or 27, wherein the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or an antigen-binding fragment thereof.

30. The method of claim 26 or 27, wherein the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an inhibitory nucleic acid selected from the group consisting of antisense polynucleotides, interfering RNA, catalytic RNA, and RNA-DNA chimeras.

31. The method of claim 26, wherein the agent that modulates CD226 expression and / or activity is an antagonist of TIGIT expression and / or activity.

32. 32. The method of claim 26 or 31, wherein the antagonist of TIGIT expression and / or activity is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

33. 32. The method of claim 26 or 31, wherein the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or an antigen-binding fragment thereof.

34. The method of claim 26 or 31, wherein the antagonist of TIGIT expression and / or activity is an inhibitory nucleic acid selected from the group consisting of an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera.

35. 27. The method of claim 26, wherein the antagonist of PVR expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

36. The method of claim 26, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVR is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

37. The method of claim 26, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

38. The method of claim 26, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

39. The method of claim 26, wherein the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

40. The method of claim 26, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

41. The method of claim 26, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

42. A method for increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that reduces or inhibits one or more additional immune co-inhibitory receptors.

43. 43. The method of claim 42, wherein the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, and CD96.

44. 43. The method of claim 42, wherein the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, and TIM3.

45. A method for increasing, enhancing or stimulating an immune response or function in an individual, comprising administering to the individual an effective amount of an agent that reduces or inhibits TIGIT expression and / or activity and an agent that increases or activates one or more additional immune co-stimulatory receptors.

46. 46. ​​The method of claim 45, wherein the one or more additional immune costimulatory receptors are selected from the group consisting of CD226, OX-40, CD28, CD27, CD137, HVEM, GITR, MICA, ICOS, NKG2D, and 2B4.

47. 46. ​​The method of claim 45, wherein the one or more additional immune co-stimulatory receptors are selected from the group consisting of CD226, OX-40, CD27, CD137, HVEM, and GITR.

48. 46. ​​The method of claim 45, wherein the one or more additional immune co-stimulatory receptors are selected from the group consisting of OX-40 and CD27.

49. 49. The method of any one of claims 1 to 48, further comprising administering at least one chemotherapeutic agent.

50. 50. The method of any one of claims 1 to 49, wherein the individual is suffering from cancer.

51. 51. The method of any one of claims 1 to 50, wherein the individual's CD4 and / or CD8 T cells have increased or enhanced priming, activation, proliferation, cytokine release and / or cytolytic activity relative to before administration of the combination.

52. 52. The method of any one of claims 1 to 51, wherein the number of CD4 and / or CD8 T cells is increased relative to before administration of the combination.

53. 53. The method of any one of claims 1 to 52, wherein the number of activated CD4 and / or CD8 T cells is increased relative to before administration of the combination.

54. Activated CD4 and / or CD8 T cells respond to γ-IFN ＋ 54. The method of any one of claims 1 to 53, characterized by an increased cytolytic activity compared to before administration of the generated CD4 and / or CD8 T cells and / or combination.

55. 55. The method of any one of claims 51 to 54, wherein the CD4 and / or CD8 T cells exhibit increased release of cytokines selected from the group consisting of IFN-γ, TNF-α and interleukins.

56. 56. The method of any one of claims 51 to 55, wherein the CD4 and / or CD8 T cells are effector memory T cells.

57. CD4 and / or CD8 effector memory T cells respond to γ-IFN ＋ 57. The method of claim 56, characterized by generated CD4 and / or CD8 T cells and / or enhanced cytolytic activity.

58. CD4 and / or CD8 effector memory T cells are CD44 ｈｉｇｈ CD62L ｌｏｗ 57. The method of claim 56, characterized by having expression of:

59. 59. The method of any one of claims 1, 2, 12, 13, 23-41, and 49-58, wherein the cancer has an increased level of T cell infiltration.

60. The method of any one of claims 1 to 11 and 42 to 59, wherein the agent that reduces or inhibits TIGIT expression and / or activity is selected from the group consisting of antagonists of TIGIT expression and / or activity, antagonists of PVR expression and / or activity, agents that inhibit and / or block the interaction of TIGIT with PVR, agents that inhibit and / or block the interaction of TIGIT with PVRL2, agents that inhibit and / or block the interaction of TIGIT with PVRL3, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVR, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL2, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL3, and combinations thereof.

61. 61. The method of claim 60, wherein the antagonist of TIGIT expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

62. 61. The method of claim 60, wherein the antagonist of PVR expression and / or activity is selected from the group consisting of a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, and an inhibitory polypeptide.

63. The method of claim 60, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVR is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

64. The method of claim 60, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVRL2 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

65. The method of claim 60, wherein the agent that inhibits and / or blocks the interaction of TIGIT with PVRL3 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

66. The method of claim 60, wherein the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

67. The method of claim 60, wherein the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

68. The method of claim 60, wherein the agent that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3 is selected from the group consisting of small molecule inhibitors, inhibitory antibodies or antigen-binding fragments thereof, aptamers, inhibitory nucleic acids, and inhibitory polypeptides.

69. 62. The method of claim 60 or 61, wherein the antagonist of TIGIT expression and / or activity is an inhibitory nucleic acid selected from the group consisting of an antisense polynucleotide, an interfering RNA, a catalytic RNA, and an RNA-DNA chimera.

70. 62. The method of claim 60 or 61, wherein the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or an antigen-binding fragment thereof.

71. The method of claim 29 or 70, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR comprising an amino acid sequence selected from the amino acid sequence (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6); or the amino acid sequence (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

72. The anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody light chain is 72. The method of any one of claims 29, 70, and 71, comprising the amino acid sequence set forth in

73. The anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody heavy chain is 73. The method of any one of claims 29 and 70 to 72, comprising the amino acid sequence set forth in

74. The anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody light chain is and the antibody heavy chain comprises the amino acid sequence set forth in 72. The method of any one of claims 29, 70, and 71, comprising the amino acid sequence set forth in

75. The method of any one of claims 29 and 70 to 74, wherein the anti-TIGIT antibody or antigen-binding fragment thereof is selected from the group consisting of a humanized antibody, a chimeric antibody, a bispecific antibody, a heteroconjugate antibody, and an immunotoxin.

76. The anti-TIGIT antibody or antigen-binding fragment thereof is selected from the group consisting of: (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6); or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7).

76. The method of any one of claims 29 and 70-75, comprising at least one HVR that is at least 90% identical to an HVR set forth in any one of: GISNRFS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

77. The anti-TIGIT antibody or a fragment thereof is a light chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in 75. The method of any one of claims 72, 73, and 74, comprising a heavy chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in

78. 80. The method of any one of claims 1-41, 43, 44, and 49-77, wherein the PD-1 axis binding antagonist is selected from the group consisting of a PD-1 binding antagonist, a PD-L1 binding antagonist, and a PD-L2 binding antagonist.

79. 79. The method of claim 78, wherein the PD-1 axis binding antagonist is a PD-1 binding antagonist.

80. 80. The method of claim 79, wherein the PD-1 binding antagonist inhibits binding of PD-1 to its ligand binding partner.

81. 81. The method of claim 79 or 80, wherein the PD-1 binding antagonist inhibits binding of PD-1 to PD-L1.

82. 81. The method of claim 79 or 80, wherein the PD-1 binding antagonist inhibits binding of PD-1 to PD-L2.

83. The method of claim 79 or 80, wherein the PD-1 binding antagonist inhibits binding of PD-1 to both PD-L1 and PD-L2.

84. 84. The method of any one of claims 79 to 83, wherein the PD-1 binding antagonist is an antibody.

85. 85. The method of any one of claims 79 to 84, wherein the PD-1 binding antagonist is MDX-1106.

86. 85. The method of any one of claims 79 to 84, wherein the PD-1 binding antagonist is MK-3475.

87. 85. The method of any one of claims 79 to 84, wherein the PD-1 binding antagonist is CT-011.

88. 84. The method of any one of claims 79 to 83, wherein the PD-1 binding antagonist is AMP-224.

89. 79. The method of claim 78, wherein the PD-1 axis binding antagonist is a PD-L1 binding antagonist.

90. 90. The method of claim 89, wherein the PD-L1 binding antagonist inhibits binding of PD-L1 to PD-1.

91. 90. The method of claim 89, wherein the PD-L1 binding antagonist inhibits binding of PD-L1 to B7-1.

92. 90. The method of claim 89, wherein the PD-L1 binding antagonist inhibits binding of PD-L1 to both PD-1 and B7-1.

93. 93. The method of any one of claims 89 to 92, wherein the PD-L1 binding antagonist is an anti-PD-L1 antibody.

94. 94. The method of any one of claims 89-93, wherein the PD-L1 binding antagonist is selected from the group consisting of YW243.55.S70, MPDL3280A, MDX-1105, and MEDI4736.

95. 94. The method of claim 93, wherein the anti-PD-L1 antibody comprises a heavy chain comprising the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO: 17), the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO: 19); and a light chain comprising the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO: 20), the HVR-L2 sequence of SASFLYS (SEQ ID NO: 21), and the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO: 22).

96. The anti-PD-L1 antibody EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23), EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 40), or EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 41) a heavy chain variable region comprising the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24) a light chain variable region comprising the amino acid sequence 94. The method of claim 93, comprising:

97. 79. The method of claim 78, wherein the PD-1 axis binding antagonist is a PD-L2 binding antagonist.

98. 98. The method of claim 97, wherein the PD-L2 binding antagonist is an antibody.

99. 98. The method of claim 97, wherein the PD-L2 binding antagonist is an immunoadhesin.

100. 99. The method of any one of claims 1, 2, 12, 13, 23-41, and 49-99, wherein the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic cancer, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and other hematological malignancies.

101. 101. The method of any one of claims 1-11 and 42-100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered continuously.

102. 101. The method of any one of claims 1-11 and 42-100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered intermittently.

103. 103. The method of any one of claims 1-11 and 49-102, wherein the agent that decreases or inhibits TIGIT expression and / or activity is administered prior to the PD-1 axis binding antagonist.

104. 103. The method of any one of claims 1-11 and 49-102, wherein the agent that decreases or inhibits TIGIT expression and / or activity is administered simultaneously with the PD-1 axis binding antagonist.

105. 103. The method of any one of claims 1-11 and 49-102, wherein the agent that decreases or inhibits TIGIT expression and / or activity is administered after the PD-1 axis binding antagonist.

106. 101. The method of any one of claims 12-41 and 49-100, wherein the PD-1 axis binding antagonist is administered prior to the agent that modulates CD226 expression and / or activity.

107. 101. The method of any one of claims 12-41 and 49-100, wherein the PD-1 axis binding antagonist is administered simultaneously with an agent that modulates CD226 expression and / or activity.

108. 101. The method of any one of claims 12-41 and 49-100, wherein the PD-1 axis binding antagonist is administered after the agent that modulates CD226 expression and / or activity.

109. 101. The method of any one of claims 42 to 44 and 49 to 100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered before the agent that reduces or inhibits one or more additional immune co-inhibitory receptors.

110. The method of any one of claims 42 to 44 and 49 to 100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered simultaneously with an agent that reduces or inhibits one or more additional immune co-inhibitory receptors.

111. The method of any one of claims 42 to 44 and 49 to 100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered after the agent that reduces or inhibits one or more additional immune co-inhibitory receptors.

112. 101. The method of any one of claims 45 to 100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered before the agent that increases or activates one or more additional immune co-stimulatory receptors.

113. 101. The method of any one of claims 45 to 100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered simultaneously with an agent that increases or activates one or more additional immune co-stimulatory receptors.

114. 101. The method of any one of claims 45 to 100, wherein the agent that reduces or inhibits TIGIT expression and / or activity is administered after the agent that increases or activates one or more additional immune co-stimulatory receptors.

115. A kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

116. A kit comprising a PD-1 axis binding antagonist, an agent that reduces or inhibits TIGIT expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

117. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with a PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

118. A kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual suffering from cancer.

119. A kit comprising a PD-1 axis binding antagonist, an agent that reduces or inhibits TIGIT expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual suffering from cancer.

120. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with a PD-1 axis binding antagonist to enhance immune function in an individual suffering from cancer.

121. A kit comprising a PD-1 axis binding antagonist and a package insert comprising instructions for using the PD-1 axis binding antagonist in combination with an agent that modulates CD226 expression and / or activity to treat or delay the progression of cancer in an individual.

122. A kit comprising a PD-1 axis binding antagonist, an agent that modulates CD226 expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that modulates CD226 expression and / or activity to treat or delay the progression of cancer in an individual.

123. A kit comprising an agent that modulates CD226 expression and / or activity and a package insert containing instructions for using the agent that modulates CD226 expression and / or activity in combination with a PD-1 axis binding antagonist to treat or delay the progression of cancer in an individual.

124. A kit comprising a PD-1 axis binding antagonist and a package insert containing instructions for using the PD-1 axis binding antagonist in combination with an agent that modulates CD226 expression and / or activity to enhance immune function in an individual suffering from cancer.

125. A kit comprising a PD-1 axis binding antagonist, an agent that modulates CD226 expression and / or activity, and a package insert containing instructions for using the PD-1 axis binding antagonist and the agent that modulates CD226 expression and / or activity to enhance immune function in an individual with cancer.

126. A kit comprising an agent that modulates CD226 expression and / or activity and a package insert containing instructions for using the agent that modulates CD226 expression and / or activity in combination with a PD-1 axis binding antagonist to enhance immune function in an individual with cancer.

127. The kit of any one of claims 115 to 126, wherein the PD-1 axis binding antagonist is an anti-PD-L1 antibody.

128. The kit of claim 127, wherein the anti-PD-L1 antibody is selected from the group consisting of YW243.55.S70, MPDL3280A, MDX-1105, and MEDI4736.

129. The kit of claim 127, wherein the anti-PD-L1 antibody comprises a heavy chain comprising the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO: 17), the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO: 18), and the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO: 19); and a light chain comprising the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO: 20), the HVR-L2 sequence of SASFLYS (SEQ ID NO: 21), and the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO: 22).

130. The anti-PD-L1 antibody EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 23), EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 40), or EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 41) a heavy chain variable region comprising the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 24) a light chain variable region comprising the amino acid sequence 128. The kit of claim 127, comprising:

131. The kit of any one of claims 115 to 126, wherein the PD-1 axis binding antagonist is an anti-PD-1 antibody.

132. The kit of claim 131, wherein the anti-PD-1 antibody is MDX-1106, MK-3475, or CT-011.

133. The kit of any one of claims 115 to 126, wherein the PD-1 axis binding antagonist is AMP-224.

134. 127. The kit of any one of claims 115 to 126, wherein the PD-1 axis binding antagonist is a PD-L2 binding antagonist.

135. 135. The kit of claim 134, wherein the PD-L2 binding antagonist is an antibody.

136. 135. The kit of claim 134, wherein the PD-L2 binding antagonist is an immunoadhesin.

137. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that reduces or inhibits one or more additional immune co-inhibitory receptors to treat or delay the progression of cancer in an individual.

138. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity, an agent that reduces or inhibits one or more additional immune co-inhibitory receptors, and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that reduces or inhibits one or more additional immune co-inhibitory receptors to treat or delay the progression of cancer in an individual.

139. A kit comprising an agent that reduces or inhibits one or more additional immune co-inhibitory receptors and a package insert containing instructions for using the agent that reduces or inhibits one or more additional immune co-inhibitory receptors in combination with an agent that reduces or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

140. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that reduces or inhibits one or more additional immune co-inhibitory receptors to enhance immune function in an individual suffering from cancer.

141. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity, an agent that reduces or inhibits one or more additional immune co-inhibitory receptors, and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that reduces or inhibits one or more additional immune co-inhibitory receptors to enhance immune function in an individual suffering from cancer.

142. A kit comprising an agent that reduces or inhibits one or more additional immune co-inhibitory receptors and a package insert containing instructions for using the agent that reduces or inhibits one or more additional immune co-inhibitory receptors in combination with an agent that reduces or inhibits TIGIT expression and / or activity to enhance immune function in an individual suffering from cancer.

143. 143. The kit of any one of claims 137 to 142, wherein the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, and CD96.

144. 143. The kit of any one of claims 137 to 142, wherein the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3 and TIM3.

145. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that increases or activates one or more additional immune co-stimulatory receptors to treat or delay the progression of cancer in an individual.

146. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity, an agent that increases or activates one or more additional immune co-stimulatory receptors, and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that increases or activates one or more additional immune co-stimulatory receptors to treat or delay the progression of cancer in an individual.

147. A kit comprising an agent that increases or activates one or more additional immune co-stimulatory receptors and a package insert containing instructions for using the agent that increases or activates one or more additional immune co-stimulatory receptors in combination with an agent that decreases or inhibits TIGIT expression and / or activity to treat or delay the progression of cancer in an individual.

148. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity in combination with an agent that increases or activates one or more additional immune co-stimulatory receptors to enhance immune function in an individual suffering from cancer.

149. A kit comprising an agent that reduces or inhibits TIGIT expression and / or activity, an agent that increases or activates one or more additional immune co-stimulatory receptors, and a package insert containing instructions for using the agent that reduces or inhibits TIGIT expression and / or activity and the agent that increases or activates one or more additional immune co-stimulatory receptors to enhance immune function in an individual suffering from cancer.

150. A kit comprising an agent that increases or activates one or more additional immune co-stimulatory receptors and a package insert containing instructions for using the agent that increases or activates one or more additional immune co-stimulatory receptors in combination with an agent that decreases or inhibits TIGIT expression and / or activity to enhance immune function in an individual suffering from cancer.

151. 151. The kit of any one of claims 145 to 150, wherein the one or more additional immune co-stimulatory receptors are selected from the group consisting of CD226, OX-40, CD28, CD27, CD137, HVEM, GITR, MICA, ICOS, NKG2D, and 2B4.

152. 151. The kit of any one of claims 145 to 150, wherein the one or more additional immune co-stimulatory receptors are selected from the group consisting of CD226, OX-40, CD27, CD137, HVEM, and GITR.

153. 151. The kit of any one of claims 145 to 150, wherein the one or more additional immune co-stimulatory receptors are selected from the group consisting of OX-40 and CD27.

154. The kit of any one of claims 115 to 120 and 127 to 153, wherein the agent that reduces or inhibits TIGIT expression and / or activity is selected from the group consisting of antagonists of TIGIT expression and / or activity, antagonists of PVR expression and / or activity, agents that inhibit and / or block the interaction of TIGIT with PVR, agents that inhibit and / or block the interaction of TIGIT with PVRL2, agents that inhibit and / or block the interaction of TIGIT with PVRL3, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVR, agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL2, and agents that inhibit and / or block intracellular signaling mediated by the binding of TIGIT to PVRL3.

155. 155. The kit of claim 154, wherein the antagonist of TIGIT expression and / or activity is an anti-TIGIT antibody or an antigen-binding fragment thereof.

156. A kit according to any one of claims 121 to 136, wherein the agent that modulates CD226 expression and / or activity is an agent that increases or stimulates CD226 expression and / or activity.

157. A kit described in any one of claims 121 to 136 and 156, wherein the agent that modulates CD226 expression and / or activity is an agent that increases and / or stimulates the interaction of CD226 with PVR.

158. A kit described in any one of claims 121 to 136, 156 and 157, wherein the agent that regulates CD226 expression and / or activity is an agent that increases and / or stimulates intracellular signaling mediated by binding of CD226 to PVR.

159. The agent that regulates CD226 expression and / or activity is an agent that inhibits and / or blocks the interaction of CD226 with TIGIT, an antagonist of TIGIT expression and / or activity, an antagonist of PVR expression and / or activity, an agent that inhibits and / or blocks the interaction of TIGIT with PVR, an agent that inhibits and / or blocks the interaction of TIGIT with PVRL2, or an agent that inhibits and / or blocks the interaction of TIGIT with PVRL3. A kit described in any one of claims 121 to 136 and 156 to 158, wherein the kit is selected from the group consisting of a drug, a drug that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVR, a drug that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL2, and a drug that inhibits and / or blocks intracellular signaling mediated by the binding of TIGIT to PVRL3.

160. The kit of claim 159, wherein the agent that regulates CD226 expression and / or activity is an agent that inhibits and / or blocks the interaction of CD226 with TIGIT.

161. The kit of claim 159 or 160, wherein the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is a small molecule inhibitor, an inhibitory antibody or antigen-binding fragment thereof, an aptamer, an inhibitory nucleic acid, or an inhibitory polypeptide.

162. The kit of claim 159 or 160, wherein the agent that inhibits and / or blocks the interaction of CD226 with TIGIT is an anti-TIGIT antibody or an antigen-binding fragment thereof.

163. The kit of claim 155 or 162, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises at least one HVR comprising an amino acid sequence selected from the amino acid sequences: (1) KSSQSLYYSGVKENLLA (SEQ ID NO: 1), ASIRFT (SEQ ID NO: 2), QQGINNPLT (SEQ ID NO: 3), GFTFSSFTMH (SEQ ID NO: 4), FIRSGSGIVFYADAVRG (SEQ ID NO: 5), and RPLGHNTFDS (SEQ ID NO: 6); or (2) RSSQSLVNSYGNTFLS (SEQ ID NO: 7), GISNRFS (SEQ ID NO: 8), LQGTHQPPT (SEQ ID NO: 9), GYSFTGHLMN (SEQ ID NO: 10), LIIPYNGGTSYNQKFKG (SEQ ID NO: 11), and GLRGFYAMDY (SEQ ID NO: 12).

164. The anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody light chain is 164. The kit of any one of claims 155, 162, and 163, comprising an amino acid sequence as set forth in

165. The anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody heavy chain is 165. A kit according to any one of claims 155 and 162 to 164, comprising an amino acid sequence as set forth in

166. The anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody light chain is and the antibody heavy chain comprises the amino acid sequence set forth in 164. The kit of any one of claims 155, 162, and 163, comprising an amino acid sequence as set forth in

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