Abstract
Hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) is mediated by an inappropriate attack by HBV-specific T cells in patients. However, this immunopathogenic process has not been clarified because of the lack of a suitable animal model. Here, we used immunocompetent Fah−/− mice as the recipients in the adoptive transfer of HBsAg+ hepatocytes from HBs-Tg mice to replace the recipient hepatocytes (HBs-HepR). HBs-HepR mice exhibited persistent HBsAg expression with chronic hepatitis and eventually developed HCC with a prevalence of 100%. HBsAg-specific CD8+ T cells were generated and specifically and continuously induced hepatocyte apoptosis with progressive chronic inflammation, and the depletion of CD8+ T cells or their deficiency prevented HCC, which could then be reproduced by the transfer of HBsAg-specific CD8+ T cells. In summary, our results demonstrated that CD8+ T cells plays a critical role in HBsAg-driven inflammtion and HCC tumorigenesis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 digital issues and online access to articles
$119.00 per year
only $9.92 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
Raw sequencing data have been deposited in GEO Datasets with the GEO accession number GSE130880.
References
Bray, F. et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 68, 394–424 (2018).
El-Serag, H. B. & Rudolph, K. L. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 132, 2557–2576 (2007).
Venook, A. P., Papandreou, C., Furuse, J. & de Guevara, L. L. The incidence and epidemiology of hepatocellular carcinoma: a global and regional perspective. Oncologist 15(Suppl. 4), 5–13 (2010).
Sung, W. K. et al. Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat. Genet. 44, 765–769 (2012).
Amaddeo, G. et al. Integration of tumour and viral genomic characterizations in HBV-related hepatocellular carcinomas. Gut 64, 820–829 (2015).
Guidotti, L. G. et al. Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. Immunity 4, 25–36 (1996).
Ando, K. et al. Mechanisms of class I restricted immunopathology. A transgenic mouse model of fulminant hepatitis. J. Exp. Med. 178, 1541–1554 (1993).
Maini, M. K. et al. The role of virus-specific CD8(+) cells in liver damage and viral control during persistent hepatitis B virus infection. J. Exp. Med. 191, 1269–1280 (2000).
Nakamoto, Y., Guidotti, L. G., Kuhlen, C. V., Fowler, P. & Chisari, F. V. Immune pathogenesis of hepatocellular carcinoma. J. Exp. Med. 188, 341–350 (1998).
Bertoletti, A. & Ferrari, C. Adaptive immunity in HBV infection. J. Hepatol. 64, S71–S83 (2016).
Guidotti, L. G. et al. Viral clearance without destruction of infected cells during acute HBV infection. Science 284, 825–829 (1999).
Guidotti, L. G. & Chisari, F. V. Noncytolytic control of viral infections by the innate and adaptive immune response. Annu. Rev. Immunol. 19, 65–91 (2001).
Thimme, R. et al. CD8(+) T cells mediate viral clearance and disease pathogenesis during acute hepatitis B virus infection. J. Virol. 77, 68–76 (2003).
Mason, W. S. et al. HBV DNA integration and clonal hepatocyte expansion in chronic hepatitis B patients considered immune tolerant. Gastroenterology 151, 986–998, e984 (2016).
Bertoletti, A., Kennedy, P. T. F. & Durantel, D. HBV infection and HCC: the ‘dangerous liaisons’. Gut 67, 787–788 (2018).
Zoulim, F. & Mason, W. S. Reasons to consider earlier treatment of chronic HBV infections. Gut 61, 333–336 (2012).
Guidotti, L. G. & Chisari, F. V. Immunobiology and pathogenesis of viral hepatitis. Annu. Rev. Pathol. 1, 23–61 (2006).
Lim, C. J. et al. Multidimensional analyses reveal distinct immune microenvironment in hepatitis B virus-related hepatocellular carcinoma. Gut 68, 916–927 (2019).
Park, J. J. et al. Hepatitis B virus-specific and global T-cell dysfunction in chronic hepatitis B. Gastroenterology 150, 684–695, e685 (2016).
Boni, C. et al. Characterization of hepatitis B virus (HBV)-specific T-cell dysfunction in chronic HBV infection. J. Virol. 81, 4215–4225 (2007).
Fisicaro, P. et al. Antiviral intrahepatic T-cell responses can be restored by blocking programmed death-1 pathway in chronic hepatitis B. Gastroenterology 138, 682–693, 693 e681–684 (2010).
Reignat, S. et al. Escaping high viral load exhaustion: CD8 cells with altered tetramer binding in chronic hepatitis B virus infection. J. Exp. Med. 195, 1089–1101 (2002).
Schuch, A. et al. Phenotypic and functional differences of HBV core-specific versus HBV polymerase-specific CD8+ T cells in chronically HBV-infected patients with low viral load. Gut 68, 905–915 (2019).
Hoogeveen, R. C. et al. Phenotype and function of HBV-specific T cells is determined by the targeted epitope in addition to the stage of infection. Gut 68, 893–904 (2019).
Cheng, Y. et al. Multifactorial heterogeneity of virus-specific T cells and association with the progression of human chronic hepatitis B infection. Sci. Immunol. 4, eaau6905 (2019).
Kim, G. A. et al. High risk of hepatocellular carcinoma and death in patients with immune-tolerant-phase chronic hepatitis B. Gut 67, 945–952 (2018).
Webster, G. J. et al. Longitudinal analysis of CD8+ T cells specific for structural and nonstructural hepatitis B virus proteins in patients with chronic hepatitis B: implications for immunotherapy. J. Virol. 78, 5707–5719 (2004).
Sitia, G. et al. Antiplatelet therapy prevents hepatocellular carcinoma and improves survival in a mouse model of chronic hepatitis B. Proc. Natl Acad. Sci. USA 109, E2165–E2172 (2012).
Sun, H., Sun, C., Xiao, W. & Sun, R. Tissue-resident lymphocytes: from adaptive to innate immunity. Cell. Mol. Immunol. 16, 205–215 (2019).
Guidotti, L. G., Matzke, B., Schaller, H. & Chisari, F. V. High-level hepatitis B virus replication in transgenic mice. J. Virol. 69, 6158–6169 (1995).
Chisari, F. V. et al. Expression of hepatitis B virus large envelope polypeptide inhibits hepatitis B surface antigen secretion in transgenic mice. J. Virol. 60, 880–887 (1986).
Isogawa, M., Chung, J., Murata, Y., Kakimi, K. & Chisari, F. V. CD40 activation rescues antiviral CD8(+) T cells from PD-1-mediated exhaustion. PLoS Pathog. 9, e1003490 (2013).
Nakamoto, Y. et al. Prevention of hepatocellular carcinoma development associated with chronic hepatitis by anti-fas ligand antibody therapy. J. Exp. Med. 196, 1105–1111 (2002).
Huang, L. R. et al. Transfer of HBV genomes using low doses of adenovirus vectors leads to persistent infection in immune competent mice. Gastroenterology 142, 1447–1450, e1443 (2012).
Huang, L. R., Wu, H. L., Chen, P. J. & Chen, D. S. An immunocompetent mouse model for the tolerance of human chronic hepatitis B virus infection. Proc. Natl Acad. Sci. USA 103, 17862–17867 (2006).
Yang, P. L., Althage, A., Chung, J. & Chisari, F. V. Hydrodynamic injection of viral DNA: a mouse model of acute hepatitis B virus infection. Proc. Natl Acad. Sci. USA 99, 13825–13830 (2002).
Brown, Z. J., Heinrich, B. & Greten, T. F. Mouse models of hepatocellular carcinoma: an overview and highlights for immunotherapy research. Nat. Rev. Gastroenterol. Hepatol. 15, 536–554 (2018).
Azuma, H. et al. Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice. Nat. Biotechnol. 25, 903–910 (2007).
Wong, M. C. S. et al. The changing epidemiology of liver diseases in the Asia-Pacific region. Nat. Rev. Gastroenterol. Hepatol. 16, 57–73 (2019).
Rehermann, B. & Thimme, R. Insights from antiviral therapy into immune responses to hepatitis B and C virus infection. Gastroenterology 156, 369–383 (2019).
Gehring, A. J. & Protzer, U. Targeting innate and adaptive immune responses to cure chronic HBV infection. Gastroenterology 156, 325–337 (2019).
Halpern, K. B. et al. Single-cell spatial reconstruction reveals global division of labour in the mammalian liver. Nature 542, 352–356 (2017).
Isogawa, M., Furuichi, Y. & Chisari, F. V. Oscillating CD8(+) T cell effector functions after antigen recognition in the liver. Immunity 23, 53–63 (2005).
Guidotti, L. G. et al. Immunosurveillance of the liver by intravascular effector CD8(+) T cells. Cell 161, 486–500 (2015).
Zong, L. et al. Breakdown of adaptive immunotolerance induces hepatocellular carcinoma in HBsAg-tg mice. Nat. Commun. 10, 221 (2019).
Jaruga, B., Hong, F., Kim, W. H. & Gao, B. IFN-gamma/STAT1 acts as a proinflammatory signal in T cell-mediated hepatitis via induction of multiple chemokines and adhesion molecules: a critical role of IRF-1. Am. J. Physiol. Gastrointest. Liver Physiol. 287, G1044–G1052 (2004).
Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411–420 (2018).
Qiu, X. et al. Single-cell mRNA quantification and differential analysis with Census. Nat. Methods 14, 309–315 (2017).
Acknowledgements
We thank Dr. Zhexiong Lian for providing the CD8 KO mice and Dr. Xin Wang for providing the Fah KO mice. We thank the NIH tetramer core facility for kindly providing HBsAg-Tetramer used in this study. This work was supported by the Chinese Academy of Science (XDB29030201), the National Key R&D Program of China (2018YFA0507403, 2017ZX10202203-009-002), and the Natural Science Foundation of China (#81788101, 81671554, 91542000, 81821001).
Author information
Authors and Affiliations
Contributions
X.H., Y.C., L.B., R.S., and Z.T. initiated and designed the research. X.H., Y.C., R.S., and Z.T. wrote the manuscript. X.H. and L.B. performed all the experiments and analyzed and interpreted the results. H.W. contributed to the discussion of the results.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Supplementary information
Rights and permissions
About this article
Cite this article
Hao, X., Chen, Y., Bai, L. et al. HBsAg-specific CD8+ T cells as an indispensable trigger to induce murine hepatocellular carcinoma. Cell Mol Immunol 18, 128–137 (2021). https://doi.org/10.1038/s41423-019-0330-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41423-019-0330-1
Keywords
This article is cited by
-
An overview of mouse models of hepatocellular carcinoma
Infectious Agents and Cancer (2023)
-
Analysis of angiogenesis-related subtypes of hepatocellular carcinoma and tumor microenvironment infiltration feature in hepatocellular carcinoma
Clinical and Translational Oncology (2023)
-
Mesencephalic astrocyte-derived neurotrophic factor reprograms macrophages to ameliorate acetaminophen-induced acute liver injury via p38 MAPK pathway
Cell Death & Disease (2022)