{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T12:21:11Z","timestamp":1768825271616,"version":"3.49.0"},"reference-count":145,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2020,5,25]],"date-time":"2020-05-25T00:00:00Z","timestamp":1590364800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Recent advancement in the immunological understanding of genesis of hepatocellular carcinoma (HCC) has implicated a decline in anti-tumour immunity on the background of chronic inflammatory state of liver parenchyma. The development of HCC involves a network of immunological activity in the tumour microenvironment involving continuous interaction between tumour and stromal cells. The reduction in anti-tumour immunity is secondary to changes in various immune cells and cytokines, and the tumour microenvironment plays a critical role in modulating the process of liver fibrosis, hepatocarcinogenesis, epithelial-mesenchymal transition (EMT), tumor invasion and metastasis. Thus, it is considered as one of primary factor behind the despicable tumour behavior and observed poor survival; along with increased risk of recurrence following treatment in HCC. The primary intent of the present review is to facilitate the understanding of the complex network of immunological interactions of various immune cells, cytokines and tumour cells associated with the development and progression of HCC.<\/jats:p>","DOI":"10.3390\/diagnostics10050338","type":"journal-article","created":{"date-parts":[[2020,5,25]],"date-time":"2020-05-25T11:42:02Z","timestamp":1590406922000},"page":"338","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Recent Advances: The Imbalance of Immune Cells and Cytokines in the Pathogenesis of Hepatocellular Carcinoma"],"prefix":"10.3390","volume":"10","author":[{"given":"Kumar","family":"Jayant","sequence":"first","affiliation":[{"name":"Warwick Medical School, University of Warwick, Coventry CV4 7H, UK"},{"name":"Department of Surgery and Cancer, Imperial College London, London SW7 5NH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nagy","family":"Habib","sequence":"additional","affiliation":[{"name":"Department of Surgery and Cancer, Imperial College London, London SW7 5NH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kai W.","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Surgery and Cancer, Imperial College London, London SW7 5NH, UK"},{"name":"Department of Surgery & Hepatitis Research Center, National Taiwan University Hospital, Taipei 10002, Taiwan"},{"name":"Centre of Mini-invasive Interventional Oncology, National Taiwan University Hospital, Taipei 10002, Taiwan"},{"name":"Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei 10002, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jane","family":"Warwick","sequence":"additional","affiliation":[{"name":"Warwick Medical School, University of Warwick, Coventry CV4 7H, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2231-3062","authenticated-orcid":false,"given":"Ramesh","family":"Arasaradnam","sequence":"additional","affiliation":[{"name":"Warwick Medical School, University of Warwick, Coventry CV4 7H, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"394","DOI":"10.3322\/caac.21492","article-title":"Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"68","author":"Bray","year":"2018","journal-title":"CA A Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S115","DOI":"10.1002\/lt.20034","article-title":"The Barcelona approach: Diagnosis, staging, and treatment of hepatocellular carcinoma","volume":"10","author":"Llovet","year":"2004","journal-title":"Liver Transplant."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1002\/hep.29086","article-title":"AASLD guidelines for the treatment of hepatocellular carcinoma","volume":"67","author":"Heimbach","year":"2017","journal-title":"Hepatology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"18059","DOI":"10.3748\/wjg.v20.i47.18059","article-title":"Hepatocellular carcinoma review: Current treatment, and evidence-based medicine","volume":"20","author":"Karaman","year":"2014","journal-title":"World J. Gastroenterol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"830","DOI":"10.1016\/j.jhep.2010.10.013","article-title":"Potential of immunotherapy for hepatocellular carcinoma","volume":"54","author":"Breous","year":"2011","journal-title":"J. Hepatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1159\/000481245","article-title":"Immuno-Oncology in Hepatocellular Carcinoma: 2017 Update","volume":"93","author":"Kudo","year":"2017","journal-title":"Oncology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1634\/theoncologist.2010-S4-14","article-title":"The Etiology of Hepatocellular Carcinoma and Consequences for Treatment","volume":"15","author":"Sanyal","year":"2010","journal-title":"Oncologist"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4103\/jcar.JCar_9_16","article-title":"Review of hepatocellular carcinoma: Epidemiology, etiology, and carcinogenesis","volume":"16","author":"Ghouri","year":"2017","journal-title":"J. Carcinog."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.metabol.2017.04.011","article-title":"Non-alcoholic fatty liver disease: A sign of systemic disease","volume":"72","author":"Reccia","year":"2017","journal-title":"Metab. Clin. Exp."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1038\/s41590-018-0044-z","article-title":"The immunology of hepatocellular carcinoma review-article","volume":"19","author":"Ringelhan","year":"2018","journal-title":"Nat. Immunol."},{"key":"ref_11","first-page":"311","article-title":"Molecular signatures and prognosis of hepatocellular carcinoma","volume":"57","author":"Hoshida","year":"2011","journal-title":"Minerva Gastroenterol. Dietol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1053\/j.gastro.2016.11.048","article-title":"Liver Cancer Cell of Origin, Molecular Class, and Effects on Patient Prognosis","volume":"152","author":"Sia","year":"2017","journal-title":"Gastroenterology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1016\/j.jhep.2011.09.020","article-title":"Obesity, inflammation, and liver cancer","volume":"56","author":"Sun","year":"2011","journal-title":"J. Hepatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1053\/j.gastro.2013.01.002","article-title":"Role of the microenvironment in the pathogenesis and treatment of hepatocellular carcinoma","volume":"144","author":"Toffanin","year":"2013","journal-title":"Gastroenterology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2013\/187204","article-title":"The Inflammatory Microenvironment in Hepatocellular Carcinoma: A Pivotal Role for Tumor-Associated Macrophages","volume":"2013","author":"Capece","year":"2012","journal-title":"BioMed Res. Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1038\/35077246","article-title":"Progress in human tumour immunology and immunotherapy","volume":"411","author":"Rosenberg","year":"2001","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1097\/CMR.0b013e32832eabd5","article-title":"Spontaneous regression of metastases from melanoma: Review of the literature","volume":"19","author":"Kalialis","year":"2009","journal-title":"Melanoma Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.ejso.2013.09.026","article-title":"Spontaneous regression of metastatic melanoma\u2014Clinical evidence of the abscopal effect","volume":"40","author":"Bramhall","year":"2014","journal-title":"Eur. J. Surg. Oncol. (EJSO)"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2557","DOI":"10.1053\/j.gastro.2007.04.061","article-title":"Hepatocellular Carcinoma: Epidemiology and Molecular Carcinogenesis","volume":"132","author":"Rudolph","year":"2007","journal-title":"Gastroenterology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.immuni.2013.07.012","article-title":"Oncology Meets Immunology: The Cancer-Immunity Cycle","volume":"39","author":"Chen","year":"2013","journal-title":"Immunity"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1159\/000449340","article-title":"Molecular Pathogenesis of Hepatocellular Carcinoma","volume":"5","author":"Ho","year":"2016","journal-title":"Liver Cancer"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1007\/s00204-012-0931-2","article-title":"Cellular and molecular mechanisms of hepatocellular carcinoma: An update","volume":"87","author":"Aravalli","year":"2012","journal-title":"Arch. Toxicol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1136\/gutjnl-2013-306627","article-title":"Hepatocellular carcinoma: Clinical frontiers and perspectives","volume":"63","author":"Bruix","year":"2014","journal-title":"Gut"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1016\/S0140-6736(18)30010-2","article-title":"Hepatocellular carcinoma","volume":"391","author":"Forner","year":"2018","journal-title":"Lancet"},{"key":"ref_25","first-page":"1","article-title":"Angiogenesis: From Chronic Liver Inflammation to Hepatocellular Carcinoma","volume":"2010","author":"Chaparro","year":"2010","journal-title":"J. Oncology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1053\/j.gastro.2017.06.007","article-title":"Identification of an Immune-specific Class of Hepatocellular Carcinoma, Based on Molecular Features","volume":"153","author":"Sia","year":"2017","journal-title":"Gastroenterology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1136\/gutjnl-2014-307990","article-title":"Current concepts of immune based treatments for patients with HCC: From basic science to novel treatment approaches","volume":"64","author":"Greten","year":"2015","journal-title":"Gut"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1046\/j.0818-9641.2001.01058.x","article-title":"Immunobiology of liver dendritic cells","volume":"80","author":"Thomson","year":"2002","journal-title":"Immunol. Cell Boil."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1136\/gut.52.2.307","article-title":"Dendritic cells and immune regulation in the liver","volume":"52","author":"Lau","year":"2003","journal-title":"Gut"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/B978-0-12-386931-9.00018-0","article-title":"Toll-IL-1-Receptor-Containing Adaptor Molecule-1: A signaling adaptor linking innate immunity to adaptive immunity","volume":"117","author":"Matsumoto","year":"2013","journal-title":"Prog. Mol. Biol. Transl. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1159\/000334245","article-title":"Dendritic Cells and Damage-Associated Molecular Patterns: Endogenous Danger Signals Linking Innate and Adaptive Immunity","volume":"4","author":"Nace","year":"2012","journal-title":"J. Innate Immun."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Khochenkov, D.A. (2008). Biology of dendritic cells. Biochem. Moscow Suppl. Ser. A, 296\u2013311.","DOI":"10.1134\/S1990747808040028"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/nri3254","article-title":"Cross-presentation by dendritic cells","volume":"12","author":"Joffre","year":"2012","journal-title":"Nat. Rev. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1038\/nri1248","article-title":"CD28-mediated co-stimulation: A quantitative support for TCR signalling","volume":"3","author":"Acuto","year":"2003","journal-title":"Nat. Rev. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1038\/nri3405","article-title":"Molecular mechanisms of T cell co-stimulation and co-inhibition","volume":"13","author":"Chen","year":"2013","journal-title":"Nat. Rev. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"603","DOI":"10.3389\/fimmu.2014.00603","article-title":"Th1\/Th2 paradigm extended: Macrophage polarization as an unappreciated pathogen-driven escape mechanism?","volume":"5","author":"Muraille","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.1158\/2159-8290.CD-16-0329","article-title":"Polarization of Tissue-Resident TFH-Like Cells in Human Hepatoma Bridges Innate Monocyte Inflammation and M2b Macrophage Polarization","volume":"6","author":"Chen","year":"2016","journal-title":"Cancer Discov."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1038\/35100512","article-title":"Cross-presentation in viral immunity and self-tolerance","volume":"1","author":"Heath","year":"2001","journal-title":"Nat. Rev. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1038\/ni1082","article-title":"Cellular mechanisms governing cross-presentation of exogenous antigens","volume":"5","author":"Ackerman","year":"2004","journal-title":"Nat. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/S0952-7915(98)80227-6","article-title":"How do cytotoxic lymphocytes kill their targets?","volume":"10","author":"Shresta","year":"1998","journal-title":"Curr. Opin. Immunol."},{"key":"ref_41","first-page":"17","article-title":"Killing mechanisms of cytotoxic T lymphocytes","volume":"13","author":"Groscurth","year":"1998","journal-title":"News Physiol. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1038\/cmi.2016.4","article-title":"Adaptive immunity in the liver","volume":"13","author":"Shuai","year":"2016","journal-title":"Cell. Mol. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1908","DOI":"10.1053\/j.gastro.2008.02.091","article-title":"Neither Multiple Tumors Nor Portal Hypertension Are Surgical Contraindications for Hepatocellular Carcinoma","volume":"134","author":"Ishizawa","year":"2008","journal-title":"Gastroenterology"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1002\/hep.24680","article-title":"Benefit of initial resection of hepatocellular carcinoma followed by transplantation in case of recurrence: An intention-to-treat analysis","volume":"55","author":"Fuks","year":"2011","journal-title":"Hepatology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1038\/nrgastro.2015.200","article-title":"Immunology in the liver\u2014From homeostasis to disease","volume":"13","author":"Heymann","year":"2016","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1038\/nrc.2016.154","article-title":"Targeting neoantigens to augment antitumour immunity","volume":"17","author":"Yarchoan","year":"2017","journal-title":"Nat. Rev. Cancer"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1126\/science.aaa4971","article-title":"Realising the promise: Neoantigens in cancer immunotherapy","volume":"348","author":"Schumacher","year":"2015","journal-title":"Science"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3413","DOI":"10.1172\/JCI80008","article-title":"Tumor neoantigens: Building a framework for personalized cancer immunotherapy","volume":"125","author":"Gubin","year":"2015","journal-title":"J. Clin. Investig."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/bs.ai.2016.01.001","article-title":"The Role of Neoantigens in Naturally Occurring and Therapeutically Induced Immune Responses to Cancer","volume":"130","author":"Ward","year":"2016","journal-title":"Adv. Immunol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1848","DOI":"10.3389\/fimmu.2017.01848","article-title":"Neoantigen targeting\u2014Dawn of a new era in cancer immunotherapy?","volume":"8","author":"Wirth","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_51","first-page":"285","article-title":"Definition of tumor-associated antigens in hepatocellular carcinoma","volume":"9","author":"Stenner","year":"2000","journal-title":"Cancer Epidemiol. Biomark. Prev."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/2162-3619-2-15","article-title":"Autoantibodies to tumor-associated antigens as biomarkers in human hepatocellular carcinoma (HCC)","volume":"2","author":"Dai","year":"2013","journal-title":"Exp. Hematol. Oncol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1102","DOI":"10.4049\/jimmunol.169.2.1102","article-title":"Large scale identification of human hepatocellular carcinoma-associated antigens by autoantibodies","volume":"169","author":"Wang","year":"2002","journal-title":"J. Immunol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1581","DOI":"10.4254\/wjh.v7.i11.1581","article-title":"Autoantibodies against tumor-associated antigens for detection of hepatocellular carcinoma","volume":"7","author":"Hong","year":"2015","journal-title":"World J. Hepatol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1111\/cas.13485","article-title":"Cancer immunotherapy-targeted glypican-3 or neoantigens","volume":"109","author":"Shimizu","year":"2018","journal-title":"Cancer Sci."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1002\/hep.26731","article-title":"Immunodominance and functional alterations of tumor-associated antigen-specific CD8+ T-cell responses in hepatocellular carcinoma","volume":"59","author":"Flecken","year":"2014","journal-title":"Hepatology"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1206","DOI":"10.1002\/hep.24149","article-title":"Comparative analysis of various tumor-associated antigen-specific t-cell responses in patients with hepatocellular carcinoma","volume":"53","author":"Mizukoshi","year":"2011","journal-title":"Hepatology"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1911","DOI":"10.1038\/bjc.2015.92","article-title":"Tumour antigen expression in hepatocellular carcinoma in a low-endemic western area","volume":"112","author":"Sideras","year":"2015","journal-title":"Br. J. Cancer"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1038\/bjc.2013.390","article-title":"Expression pattern of tumour-associated antigens in hepatocellular carcinoma: Association with immune infiltration and disease progression","volume":"109","author":"Liang","year":"2013","journal-title":"Br. J. Cancer"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Kalathil, S.G., Hutson, A., Barbi, J., Iyer, R.V., and Thanavala, Y.M. (2019). Augmentation of IFN-\u03b3+ CD8+ T cell responses correlates with survival of HCC patients on sorafenib therapy. JCI Insight, 4.","DOI":"10.1172\/jci.insight.130116"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.semcancer.2010.10.007","article-title":"The tumor microenvironment in hepatocellular carcinoma: Current status and therapeutic targets","volume":"21","author":"Yang","year":"2010","journal-title":"Semin. Cancer Boil."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1038\/nrgastro.2015.173","article-title":"Immunological landscape and immunotherapy of hepatocellular carcinoma","volume":"12","author":"Prieto","year":"2015","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/j.aohep.2019.04.003","article-title":"Immunotherapy in hepatocellular carcinoma","volume":"18","author":"Buonaguro","year":"2019","journal-title":"Ann. Hepatol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1038\/ni.1937","article-title":"Macrophage plasticity and interaction with lymphocyte subsets: Cancer as a paradigm","volume":"11","author":"Biswas","year":"2010","journal-title":"Nat. Immunol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1172\/JCI59643","article-title":"Macrophage plasticity and polarization: In vivo veritas","volume":"122","author":"Sica","year":"2012","journal-title":"J. Clin. Investig."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.immuni.2014.06.008","article-title":"Macrophage Activation and Polarization: Nomenclature and Experimental Guidelines","volume":"41","author":"Murray","year":"2014","journal-title":"Immunity"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/j.immuni.2010.05.007","article-title":"Alternative Activation of Macrophages: Mechanism and Functions","volume":"32","author":"Gordon","year":"2010","journal-title":"Immunity"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1146\/annurev.immunol.021908.132532","article-title":"Alternative Activation of Macrophages: An Immunologic Functional Perspective","volume":"27","author":"Martinez","year":"2009","journal-title":"Annu. Rev. Immunol."},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Martinez, F.O., and Gordon, S. (2014). The M1 and M2 paradigm of macrophage activation: Time for reassessment. F1000Prime Rep., 6.","DOI":"10.12703\/P6-13"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1111\/j.1365-2249.2005.02934.x","article-title":"Macrophage activation switching: An asset for the resolution of inflammation","volume":"142","author":"Porcheray","year":"2005","journal-title":"Clin. Exp. Immunol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1159\/000028096","article-title":"Alternative versus classical activation of macrophages","volume":"67","author":"Goerdt","year":"1999","journal-title":"Pathobiology"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/978-1-59745-396-7_3","article-title":"Macrophage Activation: Classical Vs. Alternative","volume":"531","author":"Classen","year":"2009","journal-title":"Methods Mol. Biol."},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Wang, N., Liang, H., and Zen, K. (2014). Molecular Mechanisms That Influence the Macrophage M1-M2 Polarization Balance. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00614"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.canlet.2019.05.019","article-title":"Individualized precision treatment: Targeting TAM in HCC","volume":"458","author":"Li","year":"2019","journal-title":"Cancer Lett."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00595-011-0058-8","article-title":"Role of tumor-associated macrophages in the progression of hepatocellular carcinoma","volume":"42","author":"Shirabe","year":"2011","journal-title":"Surg. Today"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"1646","DOI":"10.1053\/j.gastro.2016.02.040","article-title":"Tumor-Associated Neutrophils Recruit Macrophages and T-Regulatory Cells to Promote Progression of Hepatocellular Carcinoma and Resistance to Sorafenib","volume":"150","author":"Zhou","year":"2016","journal-title":"Gastroenterology"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s12935-019-0771-8","article-title":"Tumor-associated macrophages modulate resistance to oxaliplatin via inducing autophagy in hepatocellular carcinoma","volume":"19","author":"Fu","year":"2019","journal-title":"Cancer Cell Int."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1038\/nri2506","article-title":"Myeloid-derived suppressor cells as regulators of the immune system","volume":"9","author":"Gabrilovich","year":"2009","journal-title":"Nat. Rev. Immunol."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1007\/s00430-010-0151-4","article-title":"Subsets, expansion and activation of myeloid-derived suppressor cells","volume":"199","author":"Ribechini","year":"2010","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1111\/nyas.12469","article-title":"Myeloid-derived suppressor cell heterogeneity in human cancers","volume":"1319","author":"Solito","year":"2014","journal-title":"Ann. New York Acad. Sci."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1016\/j.jhep.2013.08.003","article-title":"The role of MDSCs in hepatocellular carcinoma--in vivo veritas?","volume":"59","author":"Schrader","year":"2013","journal-title":"J. Hepatol."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"43","DOI":"10.4291\/wjgp.v6.i3.43","article-title":"Emerging roles of myeloid derived suppressor cells in hepatic inflammation and fibrosis","volume":"6","author":"Hammerich","year":"2015","journal-title":"World J. Gastrointest. Pathophysiol."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1146\/annurev-med-051013-052304","article-title":"Regulation of tumor metastasis by myeloid-derived suppressor cells","volume":"66","author":"Condamine","year":"2014","journal-title":"Annu. Rev. Med."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.it.2010.10.002","article-title":"Molecular mechanisms regulating myeloid-derived suppressor cell differentiation and function","volume":"32","author":"Condamine","year":"2010","journal-title":"Trends Immunol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.4049\/jimmunol.1100164","article-title":"Activated CD69+ T Cells Foster Immune Privilege by Regulating IDO Expression in Tumor-Associated Macrophages","volume":"188","author":"Zhao","year":"2011","journal-title":"J. Immunol."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1016\/j.imbio.2013.06.010","article-title":"The role of immune-related myeloid cells in angiogenesis","volume":"218","author":"Chambers","year":"2013","journal-title":"Immunobiology"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.1172\/JCI31422","article-title":"Altered macrophage differentiation and immune dysfunction in tumor development","volume":"117","author":"Sica","year":"2007","journal-title":"J. Clin. Investig."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.ejca.2006.01.003","article-title":"Tumour-associated macrophages are a distinct M2 polarised population promoting tumour progression: Potential targets of anti-cancer therapy","volume":"42","author":"Sica","year":"2006","journal-title":"Eur. J. Cancer"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.it.2005.12.006","article-title":"The immunoregulatory role of IDO-producing human dendritic cells revisited","volume":"27","author":"Terness","year":"2006","journal-title":"Trends Immunol."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.1038\/nm934","article-title":"Evidence for a tumoral immune resistance mechanism based on tryptophan degradation by indoleamine 2,3-dioxygenase","volume":"9","author":"Uyttenhove","year":"2003","journal-title":"Nat. Med."},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"Shibata, Y., Hara, T., Nagano, J., Nakamura, N., Ohno, T., Ninomiya, S., Ito, H., Tanaka, T., Saito, K., and Seishima, M. (2016). The Role of Indoleamine 2,3-Dioxygenase in Diethylnitrosamine-Induced Liver Carcinogenesis. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0146279"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1007\/s00432-008-0395-1","article-title":"Expression and prognosis role of indoleamine 2,3-dioxygenase in hepatocellular carcinoma","volume":"134","author":"Pan","year":"2008","journal-title":"J.Cancer Res. Clin. Oncol."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"423","DOI":"10.3389\/fonc.2018.00423","article-title":"Updates in the Clinical Development of Epacadostat and Other Indoleamine 2,3-Dioxygenase 1 Inhibitors (IDO1) for Human Cancers","volume":"8","author":"Komiya","year":"2018","journal-title":"Front. Oncol."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1038\/32588","article-title":"Dendritic cells and the control of immunity","volume":"392","author":"Steinman","year":"1998","journal-title":"Nature"},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1111\/j.1600-065X.2008.00718.x","article-title":"Inflammatory signals in dendritic cell activation and the induction of adaptive immunity","volume":"227","author":"Joffre","year":"2009","journal-title":"Immunol. Rev."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"1617","DOI":"10.1002\/eji.200940153","article-title":"CD8\u03b1+ DC are not the sole subset cross-presenting cell-associated tumor antigens from a solid tumor","volume":"40","author":"McDonnell","year":"2010","journal-title":"Eur. J. Immunol."},{"key":"ref_97","unstructured":"Cancer Genome Atlas Research Network (2017). Comprehensive and Integrative Genomic Characterization of Hepatocellular Carcinoma. Cell, 169, 1327\u20131341.e23."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1038\/s41571-018-0073-4","article-title":"Molecular therapies and precision medicine for hepatocellular carcinoma","volume":"15","author":"Llovet","year":"2018","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"1972","DOI":"10.1053\/j.gastro.2008.08.008","article-title":"Pivotal role of mTOR signaling in hepatocellular carcinoma","volume":"135","author":"Villanueva","year":"2008","journal-title":"Gastroenterology"},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"1920","DOI":"10.1002\/hep.29360","article-title":"Programmed cell death-1 (PD-1) checkpoint blockade in combination with a mammalian target of rapamycin inhibitor restrains hepatocellular carcinoma growth induced by hepatoma cell-intrinsic PD-1","volume":"66","author":"Li","year":"2017","journal-title":"Hepatology"},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"e26468","DOI":"10.4161\/onci.26468","article-title":"Role of lymphocytes in liver cancer","volume":"2","author":"Mossanen","year":"2013","journal-title":"OncoImmunology"},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1016\/j.immuni.2009.05.001","article-title":"Plasticity of CD4+ T Cell Lineage Differentiation","volume":"30","author":"Zhou","year":"2009","journal-title":"Immunity"},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1189\/jlb.1107782","article-title":"Pivotal Advance: Th-1 cytokines inhibit, and Th-2 cytokines promote fibrocyte differentiation","volume":"83","author":"Shao","year":"2008","journal-title":"J. Leukoc. Biol."},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.ccr.2006.06.016","article-title":"Prediction of venous metastases, recurrence, and prognosis in hepatocellular carcinoma based on a unique immune response signature of the liver microenvironment","volume":"10","author":"Budhu","year":"2006","journal-title":"Cancer Cell"},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"2707","DOI":"10.1200\/JCO.2007.15.6521","article-title":"High Expression of Macrophage Colony-Stimulating Factor in Peritumoral Liver Tissue Is Associated With Poor Survival After Curative Resection of Hepatocellular Carcinoma","volume":"26","author":"Zhu","year":"2008","journal-title":"J. Clin. Oncol."},{"key":"ref_106","first-page":"8715","article-title":"Changes of Th1\/Th2 cytokines in patients with primary hepatocellular carcinoma after ultrasound-guided ablation","volume":"10","author":"Ji","year":"2017","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_107","first-page":"1","article-title":"Inflammatory cytokines and change of Th1\/Th2 balance as prognostic indicators for hepatocellular carcinoma in patients treated with transarterial chemoembolization","volume":"9","author":"Lee","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1111\/j.1600-065X.2011.01018.x","article-title":"Regulatory T cells and Foxp3","volume":"241","author":"Rudensky","year":"2011","journal-title":"Immunol. Rev."},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1038\/ni904","article-title":"Foxp3 programs the development and function of CD4+CD25+ regulatory T cells","volume":"4","author":"Fontenot","year":"2003","journal-title":"Nat. Immunol."},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1002\/hep.20644","article-title":"Compromised lymphocytes infiltrate hepatocellular carcinoma: The role of T-regulatory cells","volume":"41","author":"Unitt","year":"2005","journal-title":"Hepatology"},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"2457","DOI":"10.1158\/0008-5472.CAN-04-3232","article-title":"Increased Populations of Regulatory T Cells in Peripheral Blood of Patients with Hepatocellular Carcinoma","volume":"65","author":"Ormandy","year":"2005","journal-title":"Cancer Res."},{"key":"ref_112","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1007\/s12026-009-8124-7","article-title":"Regulatory T cells and immune tolerance to tumors","volume":"46","author":"Cao","year":"2009","journal-title":"Immunol. Res."},{"key":"ref_113","doi-asserted-by":"crossref","first-page":"2977","DOI":"10.3748\/wjg.v25.i24.2977","article-title":"Immunotherapy for hepatocellular carcinoma: Current and future","volume":"25","author":"Johnston","year":"2019","journal-title":"World J. Gastroenterol."},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.jaut.2009.07.010","article-title":"Application of tissue-specific NK and NKT cell activity for tumor immunotherapy","volume":"33","author":"Subleski","year":"2009","journal-title":"J. Autoimmun."},{"key":"ref_115","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1038\/nri3065","article-title":"Organ-specific features of natural killer cells","volume":"11","author":"Shi","year":"2011","journal-title":"Nat. Rev. Immunol."},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1172\/JCI66381","article-title":"Liver-resident NK cells confer adaptive immunity in skin-contact inflammation","volume":"123","author":"Peng","year":"2013","journal-title":"J. Clin. Investig."},{"key":"ref_117","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.jaut.2015.09.004","article-title":"Differential phenotypic and functional properties of liver-resident NK cells and mucosal ILC1s","volume":"67","author":"Tang","year":"2016","journal-title":"J. Autoimmun."},{"key":"ref_118","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1038\/cmi.2013.9","article-title":"NK cells in immunotolerant organs","volume":"10","author":"Sun","year":"2013","journal-title":"Cell. Mol. Immunol."},{"key":"ref_119","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1111\/imm.12224","article-title":"Subsets of human natural killer cells and their regulatory effects","volume":"141","author":"Fu","year":"2014","journal-title":"Immunology"},{"key":"ref_120","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.jaut.2015.08.011","article-title":"Human liver-resident CD56bright\/CD16neg NK cells are retained within hepatic sinusoids via the engagement of CCR5 and CXCR6 pathways","volume":"66","author":"Hudspeth","year":"2016","journal-title":"J. Autoimmun."},{"key":"ref_121","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1038\/ni1581","article-title":"Up on the tightrope: Natural killer cell activation and inhibition","volume":"9","author":"Lanier","year":"2008","journal-title":"Nat. Immunol."},{"key":"ref_122","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1146\/annurev-immunol-020711-075005","article-title":"Controlling natural killer cell responses: Integration of signals for activation and inhibition","volume":"31","author":"Long","year":"2013","journal-title":"Annu. Rev. Immunol."},{"key":"ref_123","doi-asserted-by":"crossref","unstructured":"Vermijlen, D., Luo, D., Froelich, C.J., Medema, J.P., Kummer, J.A., Willems, E., Braet, F., and Wisse, E. (2002). Hepatic natural killer cells exclusively kill splenic\/blood natural killer-resistant tumor cells by the perforin\/granzyme pathway. J. Leukoc. Biol., 72.","DOI":"10.1189\/jlb.72.4.668"},{"key":"ref_124","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1016\/j.clim.2008.08.012","article-title":"Functional impairment in circulating and intrahepatic NK cells and relative mechanism in hepatocellular carcinoma patients","volume":"129","author":"Cai","year":"2008","journal-title":"Clin. Immunol."},{"key":"ref_125","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.bbrc.2010.02.008","article-title":"Chronic hepatitis C viral infection reduces NK cell frequency and suppresses cytokine secretion: Reversion by anti-viral treatment","volume":"393","author":"Dessouki","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_126","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1002\/hep.21073","article-title":"Decreased NK cell frequency in chronic hepatitis C does not affectex vivo cytolytic killing","volume":"43","author":"Morishima","year":"2006","journal-title":"Hepatology"},{"key":"ref_127","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1189\/JLB.0309135","article-title":"Liver natural killer and natural killer T cells: Immunobiology and emerging roles in liver diseases","volume":"86","author":"Gao","year":"2009","journal-title":"J. Leukoc. Biol."},{"key":"ref_128","doi-asserted-by":"crossref","unstructured":"Baglieri, J., Brenner, D.A., and Kisseleva, T. (2019). The Role of Fibrosis and Liver-Associated Fibroblasts in the Pathogenesis of Hepatocellular Carcinoma. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20071723"},{"key":"ref_129","doi-asserted-by":"crossref","first-page":"10512","DOI":"10.3748\/wjg.v22.i48.10512","article-title":"Liver fibrosis and hepatic stellate cells: Etiology, pathological hallmarks and therapeutic targets","volume":"22","author":"Zhang","year":"2016","journal-title":"World J. Gastroenterol."},{"key":"ref_130","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1146\/annurev-pathol-052016-100322","article-title":"The Role of Cancer-Associated Fibroblasts and Fibrosisin Liver Cancer","volume":"12","author":"Affo","year":"2017","journal-title":"Annu. Rev. Pathol. Mech. Dis."},{"key":"ref_131","doi-asserted-by":"crossref","first-page":"17756","DOI":"10.3748\/wjg.v20.i47.17756","article-title":"Pathogenesis of alcoholic liver disease: Role of oxidative metabolism","volume":"20","author":"Ceni","year":"2014","journal-title":"World J. Gastroenterol."},{"key":"ref_132","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1002\/hep.27332","article-title":"Hepatic inflammation and fibrosis: Functional links and key pathways","volume":"61","author":"Seki","year":"2015","journal-title":"Hepatology"},{"key":"ref_133","doi-asserted-by":"crossref","first-page":"6841","DOI":"10.3748\/wjg.v22.i30.6841","article-title":"Cancer-associated fibroblasts in hepatocellular carcinoma","volume":"22","author":"Kubo","year":"2016","journal-title":"World J. Gastroenterol."},{"key":"ref_134","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.1158\/0008-5472.CAN-14-0243","article-title":"Transforming Growth factor-\u03b2 as a Therapeutic Target in Hepatocellular Carcinoma","volume":"74","author":"Giannelli","year":"2014","journal-title":"Cancer Res."},{"key":"ref_135","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1016\/j.celrep.2016.04.019","article-title":"Cancer-Associated Fibroblasts Regulate Tumor-Initiating Cell Plasticity in Hepatocellular Carcinoma through c-Met\/FRA1\/HEY1 Signaling","volume":"15","author":"Lau","year":"2016","journal-title":"Cell. Rep."},{"key":"ref_136","doi-asserted-by":"crossref","first-page":"118","DOI":"10.21037\/atm.2016.01.32","article-title":"Radiation therapy and the abscopal effect: A concept comes of age","volume":"4","author":"Ng","year":"2016","journal-title":"Ann. Transl. Med."},{"key":"ref_137","doi-asserted-by":"crossref","unstructured":"Huang, K.W., Jayant, K., Lee, P.-H., Yang, P.-C., Hsiao, C.-Y., Habib, N., and Sodergren, M. (2019). Positive Immuno-Modulation Following Radiofrequency Assisted Liver Resection in Hepatocellular Carcinoma. J. Clin. Med., 8.","DOI":"10.3390\/jcm8030385"},{"key":"ref_138","doi-asserted-by":"crossref","first-page":"428","DOI":"10.3390\/cancers10110428","article-title":"A systematic review and meta-analysis comparing liver resection with the Rf-based device habibTM-4X with the clamp-crush technique","volume":"10","author":"Jayant","year":"2018","journal-title":"Cancers"},{"key":"ref_139","doi-asserted-by":"crossref","first-page":"93644","DOI":"10.18632\/oncotarget.21271","article-title":"Impact of cavitron ultrasonic surgical aspirator (CUSA) and bipolar radiofrequency device (Habib-4X) based hepatectomy for hepatocellular carcinoma on tumour recurrence and disease-free survival","volume":"8","author":"Huang","year":"2017","journal-title":"Oncotarget"},{"key":"ref_140","doi-asserted-by":"crossref","first-page":"3187","DOI":"10.7150\/jca.25084","article-title":"Study to evaluate the immunomodulatory effects of radiofrequency ablation compared to surgical resection for liver cancer","volume":"9","author":"Mazmishvili","year":"2018","journal-title":"J. Cancer"},{"key":"ref_141","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1016\/j.ebiom.2018.12.058","article-title":"Comprehensive molecular and immunological characterization of hepatocellular carcinoma","volume":"40","author":"Shimada","year":"2019","journal-title":"EBioMedicine"},{"key":"ref_142","doi-asserted-by":"crossref","first-page":"7971","DOI":"10.3748\/wjg.v20.i24.7971","article-title":"Roles of Tregs in development of hepatocellular carcinoma: A meta-analysis","volume":"20","author":"Zhao","year":"2014","journal-title":"World J. Gastroenterol."},{"key":"ref_143","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1002\/cncr.29769","article-title":"Immunotherapy in hepatocellular carcinoma: Primed to make a difference?","volume":"122","author":"Harding","year":"2015","journal-title":"Cancer"},{"key":"ref_144","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1053\/j.gastro.2017.06.017","article-title":"Antibodies Against Immune Checkpoint Molecules Restore Functions of Tumor-Infiltrating T Cells in Hepatocellular Carcinomas","volume":"153","author":"Zhou","year":"2017","journal-title":"Gastroenteroloy"},{"key":"ref_145","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1016\/j.jhep.2016.10.029","article-title":"Tremelimumab in combination with ablation in patients with advanced hepatocellular carcinoma","volume":"66","author":"Duffy","year":"2016","journal-title":"J. Hepatol."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/10\/5\/338\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:32:18Z","timestamp":1760175138000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/10\/5\/338"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,25]]},"references-count":145,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["diagnostics10050338"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics10050338","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,25]]}}}