The Activation of HCV-specific CD8 T Cells by HCV Peptide Pulsed Huh7.5 Cells

Huh7.5 간암 세포주의 HCV 항원제시에 의한 HCV 특이 T 림프구의 활성에 관한 연구

  • Cho, Hyo-Sun (Department of Pharmacy, Duksung Women's University)
  • 조효선 (덕성여자대학교 약학대학 약학과)
  • Received : 2011.11.15
  • Accepted : 2011.12.10
  • Published : 2011.12.31


T cells play a key role in viral infection. However, in patients with chronic hepatitis C virus (HCV) infection, HCV-specific T cells are dysfunctional and impaired in the liver, which is the primary site for HCV replication. There are multiple potential mechanisms for HCV-specific T cell dysfunction including induction of immune inhibitory pathways (program death-1; PD-1, cytotoxic t lymphocyte associated antigen-4; CTLA-4) and immune tolerance induced specific for the liver. However, the interaction between hepatocytes and HCV-specific CD8 T cells has not clearly established. In this study, we confirmed huh (human hepatoma) 7.5 cells expressing HLA (human leukocyte antigen) A2 presented antigen to activate HCV-specific CD8 T cells in HLA A2-restricted manner and expression of PD-L (program death ligand) 1 on huh7.5 cells reduced HCV-specific CD8 T cell activation, suggesting an immune modulatory activity. Loss of HCV-specific tetramer responses following antigenic stimulation correlated with increased caspase-3 activity. In addition, PD-L1 on huh7.5 cells rescued HCV-specific CD8 T cells from apoptosis. Our results suggest that the interaction between PD-L1 and PD-1 can recover the function of HCV-specific CD8 T cells in the liver, which could be applied in therapy of HCV chronic infection.


  1. Bertolino, P., G.W. McCaughan, and D.G. Bowen. 2002. Role of primary intrahepatic T-cell activation in the 'liver tolerance effect'. Immunol. Cell. Biol. 80, 84-92.
  2. Binder, M., G. Kochs, R. Bartenschlager, and V. Lohmann. 2007. Hepatitis C virus escape from the interferon regulatory factor 3 pathway by a passive and active evasion strategy. Hepatology 46, 1365-1374.
  3. Chayama, K., C.N. Hayes, N. Hiraga, H. Abe, M. Tsuge, and M. Imamura. 2011. Animal model for study of human hepatitis viruses. J. Gastroenterol. Hepatol. 26, 13-18.
  4. Freeman, G.J., A.J. Long, and Y. Iwai. 2000. Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J. Exp. Med. 192, 1027-1034.
  5. Freeman, G.J., E.J. Wherry, R. Ahmed, and A.H. Sharpe. 2006. Reinvigorating exhausted HIV-specific T cells via PD-1-PD-1 ligand blockade. J. Exp. Med. 203, 2223-2227.
  6. Golden-Mason, L., B. Palmer, J. Klarquist, J.A. Mengshol, and N. Castelblanco. 2007. Upregulation of PD-1 expression on circulating and intrahepatic hepatitis C virus-specific CD8+ T cells associated with reversible immune dysfunction. J. Virol. 81, 9249-9258.
  7. Greenwald, R.J., G.J. Freeman, and A.H. Sharpe. 2005. The B7 family revisited. Annu. Rev. Immunol. 23, 515-548.
  8. Holz, L.E., A. Warren, D.G. Le Couteur, D.G. Bowen, and P. Bertolino. 2010. CD8+ T cell tolerance following antigen recognition on hepatocytes. J. Autoimmun. 34, 15-22.
  9. Jo, J., U. Aichele, N. Kersting, R. Klein, P. Aichele, E. Bisse, A.K. Sewell, H.E. Blum, R. Bartenschlager, V. Lohmann, and R. Thimme. 2009. Analysis of CD8+ T-cell-mediated inhibition of hepatitis C virus replication using a novel immunological model. Gastroenterology 136, 1391-1401.
  10. Jo, J., V. Lohmann, R. Bartenschlager, and R. Thimme. 2011. Experimental models to study the immunobiology of hepatitis C virus. J. Gen. Virol. 92, 477-493.
  11. Keril, J.B., A.J. McKeating, and M.C. Rice, 2002. Highly permissive cell lines for subgenomic and genomic hepatitis C virus RNA replication. J. Virol. 76, 13001-13014.
  12. Lauer, G.M. and B.D. Walker. 2001. Hepatitis C virus infection. New Engl. J. Med. 345, 41-52.
  13. Nakamoto, N., D.E. Kaplan, J. Coleclough, Y. Li, M.E. Valiga, and K.M. Chang. 2008. Functional restoration of HCV-specific CD8 T cells by PD-1 blockade is defined by PD-1 expression and compartmentalization. Gastroenterology 134, 1927-1937.
  14. Nakamoto, N., H. Cho, A. Shaked, K. Olthoff, M.E. Valiga, M. Kaminski, E. Gostick, D.A. Price, G.J. Freeman, E.J. Wherry, and K.M. Chang. 2009. Synergistic reversal of intrahepatic HCV-specific CD8 T cell exhaustion by combined PD-1/CTLA-4 blockade. PLoS Pathog. 5, e1000313.
  15. Pagliaro, L., A. Craxi, C. Cammaa, F. Tine, V. di Marco, L. Iacono, and P. Almasio. 1994. Interferon-alpha for chronic hepatitis C: an analysis of pretreatment clinical predictors of response. Hepatology 19, 820-828.
  16. Salomon, B. and J.A. Bluestone. 2001. Complexities of CD28/B7: CTLA-4 costimulatory pathways in autoimmunity and transplantation. Annu. Rev. Immunol. 19, 225-252.
  17. Uebelhoer, L., J.H. Han, B. Callendret, G. Mateu, N.H. Shoukry, H.L. Hanson, C.M. Rice, C.M. Walker, and A. Grakoui. 2008. Stable cytotoxic T cell escape mutation in hepatitis C virus is linked to maintenance of viral fitness. PLoS Pathog. 4, e1000143.
  18. Varela-Rohena, A., C. Carpenito, E.E. Perez, M. Richardson, R.V. Parry, M. Milone, J. Scholler, X. Hao, A. Mexas, R.G. Carroll, C.H. June, and J.L. Riley. 2008. Genetic engineering of T cells for adoptive immunotherapy. Immunol. Res. 42, 166-181.
  19. Wherry, E.J. and R. Ahmed. 2004. Memory CD8 T-cell differentiation during viral infection. J. Virol. 8, 5535-5545.
  20. Willberg, C., E. Barnes, and P. Klenerman. 2003. HCV immunology-death and the maiden T cell. Cell. Death Differ. 10, S39-47.