Reduced CD160 Expression Contributes to Impaired NK-cell Function and Poor Clinical Outcomes in Patients with HCC.

: We previously reported that deficiencies in natural killer (NK)-cell number and function play an important role in the progression of hepatocellular carcinoma (HCC). However, the mechanisms underlying this phenomenon remain obscure. In this study, we analyzed the expression of CD160 on intrahepatic NK cells by evaluating peritumoral and intratumoral tissues of 279 patients with HCC and 20 healthy livers. We observed reduced expression of CD160 on intratumoral NK cells, and patients with lower CD160 cell densities within tumor cells exhibited worse disease and a higher recurrence rate. High-resolution microarray and gene set enrichment analysis of flow cytometry-sorted primary intrahepatic CD160+ and CD160- NK cells of healthy livers indicated that human CD160+ NK cells exhibited functional activation, high IFNγ production, and NK-mediated immunity. In addition, global transcriptomic analysis of sorted peritumoral and intratumoral CD160+ NK cells revealed that intratumoral CD160+ NK cells are more exhausted than peritumoral CD160+ NK cells and produce less IFNγ. High levels of TGFβ1 interfered with production of IFNγ by CD160+ NK cells, blocking of which specifically restored IFNγ production in CD160+ NK cells to normal levels. These findings indicate that reduced numbers of CD160+ NK cells, together with the functional impairment of CD160+ NK cells by TGFβ1, contribute to tumor immune escape. In addition, restoring the expression of CD160 and blocking TGFβ1 appear a promising therapeutic strategy against liver cancer. SIGNIFICANCE: These findings show that reduced number and function of CD160+ NK cells in the tumor microenvironment contributes to immune escape of HCC; blocking TGFβ1 restores IFNγ production of CD160+ NK cells.Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/78/23/6581/F1.large.jpg.

[1]  Zonghai Li,et al.  Natural killer cells in hepatocellular carcinoma: current status and perspectives for future immunotherapeutic approaches , 2017, Frontiers of Medicine.

[2]  R. Sun,et al.  Liver-resident NK cells and their potential functions , 2017, Cellular & Molecular Immunology.

[3]  P. Chow,et al.  Delineation of an immunosuppressive gradient in hepatocellular carcinoma using high-dimensional proteomic and transcriptomic analyses , 2017, Proceedings of the National Academy of Sciences.

[4]  Meijuan Huang,et al.  High NKG2A expression contributes to NK cell exhaustion and predicts a poor prognosis of patients with liver cancer , 2017, Oncoimmunology.

[5]  W. Yin,et al.  Liver-infiltrating CD11b−CD27− NK subsets account for NK-cell dysfunction in patients with hepatocellular carcinoma and are associated with tumor progression , 2016, Cellular and Molecular Immunology.

[6]  C. O’Farrelly,et al.  Liver immunology and its role in inflammation and homeostasis , 2016, Cellular & Molecular Immunology.

[7]  A. Grakoui,et al.  Presentation of hepatocellular antigens , 2016, Cellular and Molecular Immunology.

[8]  E. Wisse,et al.  Liver natural killer cells: subsets and roles in liver immunity , 2015, Cellular and Molecular Immunology.

[9]  J. Xu,et al.  The predictive value of centre tumour CD8+ T cells in patients with hepatocellular carcinoma: comparison with Immunoscore , 2015, OncoTarget.

[10]  H. Favoreel,et al.  Role of NK cells in immunotherapy and virotherapy of solid tumors. , 2015, Immunotherapy.

[11]  P. Feugier,et al.  Combination of CD160 and CD200 as a useful tool for differential diagnosis between chronic lymphocytic leukemia and other mature B‐cell neoplasms , 2015, International journal of laboratory hematology.

[12]  Cheng-song Sun,et al.  Natural killer cell dysfunction in hepatocellular carcinoma and NK cell-based immunotherapy , 2015, Acta Pharmacologica Sinica.

[13]  Nicholas K. Brown,et al.  CD160 is essential for NK-mediated IFN-γ production , 2015, The Journal of experimental medicine.

[14]  A. Prentice,et al.  Minimal residual disease detection with tumor-specific CD160 correlates with event-free survival in chronic lymphocytic leukemia , 2015, Blood Cancer Journal.

[15]  Z. Tian,et al.  NK cell receptor imbalance and NK cell dysfunction in HBV infection and hepatocellular carcinoma , 2014, Cellular and Molecular Immunology.

[16]  A. Farina,et al.  CD160-Associated CD8 T-Cell Functional Impairment Is Independent of PD-1 Expression , 2014, PLoS pathogens.

[17]  G. Giannelli,et al.  Transforming growth factor-β as a therapeutic target in hepatocellular carcinoma. , 2014, Cancer research.

[18]  M. Carrington,et al.  Liver environment and HCV replication affect human T-cell phenotype and expression of inhibitory receptors. , 2014, Gastroenterology.

[19]  Matthew G. Macauley,et al.  CD160 Activation by Herpesvirus Entry Mediator Augments Inflammatory Cytokine Production and Cytolytic Function by NK Cells , 2013, The Journal of Immunology.

[20]  M. Colonna,et al.  Intraepithelial type 1 innate lymphoid cells are a unique subset of IL-12- and IL-15-responsive IFN-γ-producing cells. , 2013, Immunity.

[21]  Benjamin E. Gross,et al.  Integrative Analysis of Complex Cancer Genomics and Clinical Profiles Using the cBioPortal , 2013, Science Signaling.

[22]  J. Gribben,et al.  T cells from CLL patients exhibit features of T-cell exhaustion but retain capacity for cytokine production. , 2013, Blood.

[23]  Benjamin E. Gross,et al.  The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. , 2012, Cancer discovery.

[24]  M. Shiroishi,et al.  Molecular basis for herpesvirus entry mediator recognition by the human immune inhibitory receptor CD160 and its relationship to the cosignaling molecules BTLA and LIGHT. , 2011, Journal of molecular biology.

[25]  I. Ng,et al.  Chemokine-driven lymphocyte infiltration: an early intratumoural event determining long-term survival in resectable hepatocellular carcinoma , 2011, Gut.

[26]  M. Calaminici,et al.  Differential and tumor-specific expression of CD160 in B-cell malignancies. , 2011, Blood.

[27]  M. Manns,et al.  Dual Function of the NK Cell Receptor 2B4 (CD244) in the Regulation of HCV-Specific CD8+ T Cells , 2011, PLoS pathogens.

[28]  G. Giannelli,et al.  Inhibiting TGF-β signaling in hepatocellular carcinoma. , 2011, Biochimica et biophysica acta.

[29]  H. Pircher,et al.  Coexpression of PD-1, 2B4, CD160 and KLRG1 on Exhausted HCV-Specific CD8+ T Cells Is Linked to Antigen Recognition and T Cell Differentiation , 2010, PLoS pathogens.

[30]  J. Gribben,et al.  CD160 signaling mediates PI3K-dependent survival and growth signals in chronic lymphocytic leukemia. , 2010, Blood.

[31]  G. Freeman,et al.  The CD160, BTLA, LIGHT/HVEM pathway: a bidirectional switch regulating T‐cell activation , 2009, Immunological reviews.

[32]  Eric Vivier,et al.  Functions of natural killer cells , 2008, Nature Immunology.

[33]  G. Cai,et al.  Corrigendum: CD160 inhibits activation of human CD4+ T cells through interaction with herpesvirus entry mediator , 2008, Nature Immunology.

[34]  S. Valitutti,et al.  CD160-activating NK cell effector functions depend on the phosphatidylinositol 3-kinase recruitment. , 2007, International immunology.

[35]  A. Bensussan,et al.  A Soluble Form of the MHC Class I-Specific CD160 Receptor Is Released from Human Activated NK Lymphocytes and Inhibits Cell-Mediated Cytotoxicity1 , 2007, The Journal of Immunology.

[36]  Z. Trajanoski,et al.  Type, Density, and Location of Immune Cells Within Human Colorectal Tumors Predict Clinical Outcome , 2006, Science.

[37]  T. Yamamura,et al.  Murine CD160, Ig-Like Receptor on NK Cells and NKT Cells, Recognizes Classical and Nonclassical MHC Class I and Regulates NK Cell Activation1 , 2005, The Journal of Immunology.

[38]  P. Le Bouteiller,et al.  Cutting Edge: Engagement of CD160 by its HLA-C Physiological Ligand Triggers a Unique Cytokine Profile Secretion in the Cytotoxic Peripheral Blood NK Cell Subset1 , 2004, The Journal of Immunology.

[39]  G. Gerken,et al.  Increased Levels of Interleukin-10 in Serum from Patients with Hepatocellular Carcinoma Correlate with Profound Numerical Deficiencies and Immature Phenotype of Circulating Dendritic Cell Subsets , 2004, Clinical Cancer Research.

[40]  F. Mami-Chouaib,et al.  Engagement of CD160 receptor by HLA-C is a triggering mechanism used by circulating natural killer (NK) cells to mediate cytotoxicity , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[41]  M. Nikolova,et al.  BY55/CD160 acts as a co-receptor in TCR signal transduction of a human circulating cytotoxic effector T lymphocyte subset lacking CD28 expression. , 2002, International immunology.

[42]  G. Freeman,et al.  Cloning of BY55, a novel Ig superfamily member expressed on NK cells, CTL, and intestinal intraepithelial lymphocytes. , 1998, Journal of immunology.

[43]  G. Leca,et al.  A novel 80-kD cell surface structure identifies human circulating lymphocytes with natural killer activity , 1993, The Journal of experimental medicine.

[44]  Meijuan Huang,et al.  Reduced CD 160 expression contributes to impaired NK cell function and poor clinical outcomes in patients with HCC Running Title : CD 160 in NK cell functional impairment and HCC , 2018 .

[45]  S. Yasuda,et al.  Clinical impact of herpesvirus entry mediator expression in human hepatocellular carcinoma. , 2015, European journal of cancer.

[46]  A. Lechtermann,et al.  Exercise affects the gene expression profiles of human white blood cells. , 2007, Journal of applied physiology.