Tumor-infiltrating NY-ESO-1–specific CD8 ⁺ T cells are negatively regulated by LAG-3 and PD-1 in human ovarian cancer

Abstract

NY-ESO-1 is a "cancer-testis" antigen frequently expressed in epithelial ovarian cancer (EOC) and is among the most immunogenic tumor antigens defined to date. In an effort to understand in vivo tolerance mechanisms, we assessed the phenotype and function of NY-ESO-1-specific CD8 + T cells derived from peripheral blood lymphocytes (PBLs), tumor-infiltrating lymphocytes (TILs), and tumorassociated lymphocytes (TALs) of EOC patients with NY-ESO-1-expressing tumors, with or without humoral immunity to NY-ESO-1. Whereas NY-ESO-1-specific CD8 + T cells were readily detectable ex vivo with tetramers in TILs and TALs of seropositive patients, they were only detectable in PBLs following in vitro stimulation. Compared with PBLs, tumor-derived NY-ESO-1-specific CD8 + T cells demonstrated impaired effector function, preferential usage of dominant T-cell receptor, and enriched coexpression of inhibitory molecules LAG-3 and PD-1. Expression of LAG-3 and PD-1 on CD8 + T cells was up-regulated by IL-10, IL-6 (cytokines found in tumor ascites), and tumor-derived antigen-presenting cells. Functionally, CD8 + LAG-3 + PD-1 + T cells were more impaired in IFN-γ/TNF-α production compared with LAG-3 + PD-1 − or LAG-3 − PD-1 − subsets. Dual blockade of LAG-3 and PD-1 during T-cell priming efficiently augmented proliferation and cytokine production by NY-ESO-1-specific CD8 + T cells, indicating that antitumor function of NY-ESO-1-specific CD8 + T cells could potentially be improved by therapeutic targeting of these inhibitory receptors.tumor-infiltrating lymphocytes | IL-6 | IL-10 | T cell receptor T he presence of tumor-infiltrating lymphocytes within the tumor microenvironment is considered to be an indication of the host immune response to tumor antigens and is thought to reflect the dynamic process of "cancer immunoediting" (1). In epithelial ovarian cancer (EOC), support for the role of immune surveillance of tumors comes from observations indicating that the presence of intraepithelial CD8 + -infiltrating T lymphocytes in tumors is associated with improved survival of patients with the disease (2). Although several lines of evidence have shown that spontaneous or vaccine-induced tumor-antigen-specific CD8 + T cells can recognize EOC targets (3), prolongation of survival in patients treated with immunization has only rarely been observed. This is probably a reflection of several in vivo immunosuppressive mechanisms in EOC-bearing hosts (4). Therefore, understanding factors that regulate the function(s) of tumor-antigen-specific CD8 + T cells is critical for effective control of tumor recurrence.The NY-ESO-1 tumor antigen is a major target of CD8 + T cell recognition in EOC, eliciting both cellular and humoral immune responses in a proportion of patients with advanced NY-ESO-1-expressing tumors (5). However, similar to infectious disease models, chronic antigenic stimulation may result in exhaustion of antigen-specific CD8 + T cells (6) and loss of ability to produce key cytokines that are critical for the maintenance of CD8 + T ce...

PD-1 and CD103 Are Widely Coexpressed on Prognostically Favorable Intraepithelial CD8 T Cells in Human Ovarian Cancer

“…Together, these features suggest that TIL in ovarian cancer may have experienced prolonged exposure to their cognate antigen and, as a consequence, may be functionally exhausted. In support of this notion, prior studies have shown that NY-ESO-1-specific CD8 þ TIL from ovarian cancer patients frequently express PD-1 and are functionally impaired compared with their counterparts in peripheral blood (20). Moreover, ovarian tumor cells have been reported to express high levels of PD-L1 (21,22).…”

Blockade of LAG3 enhances responses of tumor-infiltrating T cells in mismatch repair-proficient liver metastases of colorectal cancer

“…TIL and PBMC from LM-CRC were labeled with 0.1 µM carboxyfluorescein diacetate succinimidyl ester (CFSE, Invitrogen); afterwards 10 5 TIL or PBMC were cultured in 200 µl complete medium in each well of a 96-well round-bottom culture plate and stimulated with anti-human CD3/CD28 dynabeads (Gibco-Life Technologies AS, Norway) at a cell: bead ratio of 10:1, in the presence or absence of 10 μg/ml antagonistic monoclonal antibodies against human PD-L1 (clone 5H1, kindly provided by Dr. Haidong Dong, Mayo Clinic College of Medicine, 59 ) TIM3 (clone F38-2E2, Biolegend, San Diego, USA, 60 , 61 ) LAG3 (clone 17B4, AdipoGen, Liestal, Switzerland 62 ) or CTLA4 (clone BNI3, Beckman Coulter, Marseille, France, 63 ) or isotype-matched control antibodies (mIgG1 and mIgG2 a, Biolegend, London, UK). In preliminary experiments a cell: bead ratio of 10:1 was established to provide sub-optimal stimulation of T cell proliferation.…”

Management of Italian Patients With Advanced Non–Small-Cell Lung Cancer After Second-Line Treatment: Results of the Longitudinal Phase of the LIFE Observational Study

“…Besides, the function of tumor antigen-specific CD8 þ T cells was significantly augmented by dual blockade of LAG3 and PD-1, thus, possibly represented a highly promising immune-based strategy in cancer treatment. 38,39 There are multiple clinical trials trying to translate combination modality into clinical practice (Table 1).…”