PMID- 28648905 OWN - NLM STAT- MEDLINE DCOM- 20171016 LR - 20221207 IS - 1528-0012 (Electronic) IS - 0016-5085 (Linking) VI - 153 IP - 4 DP - 2017 Oct TI - Antibodies Against Immune Checkpoint Molecules Restore Functions of Tumor-Infiltrating T Cells in Hepatocellular Carcinomas. PG - 1107-1119.e10 LID - S0016-5085(17)35802-X [pii] LID - 10.1053/j.gastro.2017.06.017 [doi] AB - BACKGROUND & AIMS: Ligand binding to inhibitory receptors on immune cells, such as programmed cell death 1 (PD-1) and cytotoxic T-lymphocyte associated protein 4 (CTLA4), down-regulates the T-cell-mediated immune response (called immune checkpoints). Antibodies that block these receptors increase antitumor immunity in patients with melanoma, non-small-cell lung cancer, and renal cell cancer. Tumor-infiltrating CD4(+) and CD8(+) T cells in patients with hepatocellular carcinoma (HCC) have been found to be functionally compromised. We analyzed HCC samples from patients to determine if these inhibitory pathways prevent T-cell responses in HCCs and to find ways to restore their antitumor functions. METHODS: We collected HCC samples from 59 patients who underwent surgical resection from November 2013 through May 2017, along with tumor-free liver tissues (control tissues) and peripheral blood samples. We isolated tumor-infiltrating lymphocytes (TIL) and intra-hepatic lymphocytes. We used flow cytometry to quantify expression of the inhibitory receptors PD-1, hepatitis A virus cellular receptor 2 (TIM3), lymphocyte activating 3 (LAG3), and CTLA4 on CD8(+) and CD4(+) T cells from tumor, control tissue, and blood; we studied the effects of antibodies that block these pathways in T-cell activation assays. RESULTS: Expression of PD-1, TIM3, LAG3, and CTLA4 was significantly higher on CD8(+) and CD4(+) T cells isolated from HCC tissue than control tissue or blood. Dendritic cells, monocytes, and B cells in HCC tumors expressed ligands for these receptors. Expression of PD-1, TIM3, and LAG3 was higher on tumor-associated antigen (TAA)-specific CD8(+) TIL, compared with other CD8(+) TIL. Compared with TIL that did not express these inhibitory receptors, CD8(+) and CD4(+) TIL that did express these receptors had higher levels of markers of activation, but similar or decreased levels of granzyme B and effector cytokines. Antibodies against CD274 (PD-ligand1 [PD-L1]), TIM3, or LAG3 increased proliferation of CD8(+) and CD4(+) TIL and cytokine production in response to stimulation with polyclonal antigens or TAA. Importantly, combining antibody against PD-L1 with antibodies against TIM3, LAG3, or CTLA4 further increased TIL functions. CONCLUSIONS: The immune checkpoint inhibitory molecules PD-1, TIM3, and LAG3 are up-regulated on TAA-specific T cells isolated from human HCC tissues, compared with T cells from tumor-free liver tissues or blood. Antibodies against PD-L1, TIM3, or LAG3 restore responses of HCC-derived T cells to tumor antigens, and combinations of the antibodies have additive effects. Strategies to block PD-L1, TIM3, and LAG3 might be developed for treatment of primary liver cancer. CI - Copyright (c) 2017 AGA Institute. Published by Elsevier Inc. All rights reserved. FAU - Zhou, Guoying AU - Zhou G AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Sprengers, Dave AU - Sprengers D AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Boor, Patrick P C AU - Boor PPC AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Doukas, Michail AU - Doukas M AD - Department of Pathology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Schutz, Hannah AU - Schutz H AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Mancham, Shanta AU - Mancham S AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Pedroza-Gonzalez, Alexander AU - Pedroza-Gonzalez A AD - Laboratory of Immunology Research, FES-Iztacala, UNAM, Mexico. FAU - Polak, Wojciech G AU - Polak WG AD - Department of Surgery, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - de Jonge, Jeroen AU - de Jonge J AD - Department of Surgery, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Gaspersz, Marcia AU - Gaspersz M AD - Department of Surgery, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Dong, Haidong AU - Dong H AD - Department of Urology and Immunology, Mayo Clinic College of Medicine, Rochester, Minnesota. FAU - Thielemans, Kris AU - Thielemans K AD - Laboratory of Molecular and Cellular Therapy, Department of Immunology-Physiology, Vrije Universiteit, Brussels, and eTheRNA immunotherapies NV, Niel, Belgium. FAU - Pan, Qiuwei AU - Pan Q AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - IJzermans, Jan N M AU - IJzermans JNM AD - Laboratory of Immunology Research, FES-Iztacala, UNAM, Mexico. FAU - Bruno, Marco J AU - Bruno MJ AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. FAU - Kwekkeboom, Jaap AU - Kwekkeboom J AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Centre, Rotterdam, the Netherlands. Electronic address: j.kwekkeboom@erasmusmc.nl. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170623 PL - United States TA - Gastroenterology JT - Gastroenterology JID - 0374630 RN - 0 (Antibodies, Monoclonal) RN - 0 (Antibodies, Neutralizing) RN - 0 (Antigens, CD) RN - 0 (Antineoplastic Agents) RN - 0 (CTLA-4 Antigen) RN - 0 (CTLA4 protein, human) RN - 0 (Cytokines) RN - 0 (HAVCR2 protein, human) RN - 0 (Hepatitis A Virus Cellular Receptor 2) RN - 0 (PDCD1 protein, human) RN - 0 (Programmed Cell Death 1 Receptor) RN - 0 (Lymphocyte Activation Gene 3 Protein) RN - 0 (Lag3 protein, human) SB - IM MH - Antibodies, Monoclonal/*pharmacology MH - Antibodies, Neutralizing/*pharmacology MH - *Antigens, CD/immunology/metabolism MH - Antineoplastic Agents/*pharmacology MH - Antineoplastic Combined Chemotherapy Protocols/pharmacology MH - CTLA-4 Antigen/antagonists & inhibitors/immunology/metabolism MH - Carcinoma, Hepatocellular/*drug therapy/immunology/metabolism/pathology MH - Cell Proliferation/drug effects MH - Cells, Cultured MH - Coculture Techniques MH - Cytokines/metabolism MH - Hepatitis A Virus Cellular Receptor 2/*antagonists & inhibitors/immunology/metabolism MH - Humans MH - Immunotherapy/*methods MH - Liver Neoplasms/*drug therapy/immunology/metabolism/pathology MH - Lymphocyte Activation/drug effects MH - Lymphocytes, Tumor-Infiltrating/*drug effects/immunology/metabolism MH - Programmed Cell Death 1 Receptor/*antagonists & inhibitors/immunology/metabolism MH - Signal Transduction/drug effects MH - T-Lymphocytes/*drug effects/immunology/metabolism MH - Tumor Escape/drug effects MH - Tumor Microenvironment MH - Up-Regulation MH - Lymphocyte Activation Gene 3 Protein OTO - NOTNLM OT - GPC3 OT - Galectin 9 OT - Immunotherapy OT - MAGEC2 EDAT- 2017/06/27 06:00 MHDA- 2017/10/17 06:00 CRDT- 2017/06/27 06:00 PHST- 2016/12/02 00:00 [received] PHST- 2017/06/05 00:00 [revised] PHST- 2017/06/15 00:00 [accepted] PHST- 2017/06/27 06:00 [pubmed] PHST- 2017/10/17 06:00 [medline] PHST- 2017/06/27 06:00 [entrez] AID - S0016-5085(17)35802-X [pii] AID - 10.1053/j.gastro.2017.06.017 [doi] PST - ppublish SO - Gastroenterology. 2017 Oct;153(4):1107-1119.e10. doi: 10.1053/j.gastro.2017.06.017. Epub 2017 Jun 23.