摘要
乙肝病毒(HBV)为一类嗜肝性病毒,在世界范围内广泛流行,在中国HBV感染情况更为严重。HBV感染对人类健康具有极大的威胁,也给人类社会造成重大的损失。研究认为特异性细胞毒T细胞(CTL)反应在清除HBV感染的过程中发挥着极其重要的作用。HBV感染后引起的CTL反应是一把双刃剑,既起着病毒清除的作用,但同时又造成了肝损伤。而病毒为了维护自身的生存会通过产生各种突变体来对抗机体的清除作用,且HBV突变被证明与乙肝疾病发展有着一定的关系。因此,HBV感染中CTL反应相关研究具有重要的理论及实际意义。
目前已报道的CTL表位主要是由西方学者发现的,但西方国家与亚洲国家主要流行的HBV基因型并不相同。基于这个情况,我们选取中国分布最多的B型和C型HBV作为研究对象,并选择了HBV组成成分中保守性最好且免疫原性最强的核心蛋白(HBc)作为研究目标。通过总结分析NCBI上171条B型HBc序列和159条C型HBc序列,利用overlapping方法合成涵盖HBcl-150全序列的九肽(每条多肽与相邻多肽有8个氨基酸的重复),若某一氨基酸的突变率超过10%,覆盖该突变氨基酸的一系列多肽也被包括在合成的范围内,合计共191条九肽。首先通过T2细胞结合实验,筛选到四条HLA-A2高结合力的多肽(HBc60-68*,HBc123-131,HBc123-13l*,HBc141-149).随后通过晶体结构学研究显示这四条多肽具有典型的HLA-A*0201表位的结构特点,但同时也具有一些新的特点,从而拓展了人们对MHC-多肽复合物结构特点上的认识。其中HBc第60位突变(L60V)产生的新CTL表位(HBc60-68*,V60),引起我们极大的兴趣,并对这一突变位点进行了深入的研究。通过晶体学方法和功能学实验相结合的策略,证明V60具有典型的HLA-A*0201表位多肽的结构特征,在小鼠体中可以产生很强的特异性CTL反应,而其野生型多肽L60并不可以。通过分析HBV感染患者的临床资料,观察到HBc L60V突变与临床更强的肝损伤,更高病毒水平以及不良预后相关。随后通过在HLA-A2.1/HBV杂交小鼠中进行V60多肽免疫,我们证明V60刺激起的多肽特异性CTL反应在杂交小鼠中造成了一定的肝损伤。另一方面,我们在野生型HBV质粒中引入HBc L60V突变,在质粒转染Huh7细胞后检测发现,突变型质粒的HBV复制水平明显高于野生型质粒,高压尾脉注射野生型HBV质粒与突变型质粒进入小鼠体内也得到了同样的实验结果。我们进一步通过细胞内和原核表达野生型及突变型HBc蛋白,发现在同等HBc蛋白表达量的情况下,突变型HBc蛋白包装成核心颗粒的水平要明显高于野生型蛋白,这就说明了HBc L60V突变可能通过促进HBc核衣壳的包装来促进HBV复制,而这个结果也解释了含有HBcL60V突变的慢肝病人中HBV DNA水平更高的现象。本实验中我们展示了一种新型HBV突变行为,该突变以产生一个新HLA-A2限制性CTL表位的代价而促进了HBV复制。我们的实验结果表明HBc L60V突变影响了T细胞反应,HBV复制以及艺肝疾病发展,有助于人们对临床乙肝疾病发生机制的了解。
Hepatitis B virus (HBV) is a strictly hepatotropic DNA virus and has a worldwide distribution, especially in China. As the pathogenic role of HBV, it is a great threat to the health of human and can cause huge losses to our society. Many previous reports suggest that cytotoxic T lymphocyte (CTL) response plays a critical role in clearing HBV infection. CTL response induced by HBV infection is a "double-edge sword". It can clear virus infection as well as cause liver injury. As a highly prevalent hepadnavirus, HBV can evolve by mutations to increase its adaption against environmental selection, and these mutants are closely correlated with the pathogenesis of HBV infection. Based on these results, study of CTL response exhibits important theoretical and practical significances.
Most of the reported CTL epitopes have been defined in HBV genotype A and D, lacking from genotype B and C, which is the most prevalent HBV genotype in China. In this study, the overlapping9-mer peptide pool (8amino acids overlap) covering HBV core protein (HBc)1-150and its variants was built to identify HBc of genotype B and C derived CTL epitopes. A total of191nonapeptides were included. Four peptides of high binding capacity to HLA-A2molecular were selected by MHC stabilization assay (HBc60-68*, HBc123-131, HBcl23-131*, HBc141-149) and the structures of MHC-peptide complex showed these four peptides are typical HLA-A2restricted peptides but also with their own characteristics, which may expand understandings of binding features of HLA-A2restricted epitopes. Of note, HBc60-68*(V60) generated by the L60V variation in HBc was carefully studied by structural and immunogenic analysis. Clinical researches showed that The HBc L60V variation is correlated with hepatic necroinflammation and higher viral levels, and it may be associated with poor prognosis in CHB patients. Immunization with the defined HBV epitope V60peptide elicited specific CTL-induced liver injury in HLA-A2+/HBV transgenic mice. In addition, in vitro and in vivo experiments both demonstrated that the HBc L60V variation facilitates viral capsid assembly and increases HBV replication. These data suggest that the HBc L60V variation can impact both HBV replication and HBV-specific T cell responses. Therefore, our work provides further dissection of the impact of the HBc L60V variation. which orchestrates HBV replication, viral persistence, and immunopathogenesis during chronic viral infection.
引文
1. Lee, R. C., R. L. Feinbaum, and V. Ambros.1993. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75:843-854.
2. Wightman, B., I. Ha, and G. Ruvkun.1993. Posttranscriptional regulation of the-heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans. Cell 75:855-862.
3. Brodersen, P., L. Sakvarelidze-Achard, M. Bruun-Rasmussen, P. Dunoyer, Y. Y. Yamamoto, L. Sieburth, and O. Voinnet.2008. Widespread translational inhibition by plant miRNAs and siRNAs. Science 320:1185-1190.
4. Lanet, E., E. Delannoy, R. Sormani, M. Floris, P. Brodersen, P. Crete, O. Voinnet, and C. Robaglia.2009. Biochemical Evidence for Translational Repression by Arabidopsis MicroRNAs. Plant Cell 21:1762-1768.
5. Girard, A., R. Sachidanandam, G. J. Hannon, and M. A. Carmell.2006. A germline-specific class of small RNAs binds mammalian Piwi proteins. Nature 442:199-202.
6. Gunawardane, L. S., K. Saito, K. M. Nishida, K. Miyoshi, Y. Kawamura, T. Nagami, H. Siomi, and M. C. Siomi.2007. A slicer-mediated mechanism for repeat-associated siRNA 5'end formation in Drosophila. Science 315:1587-1590.
7. Jopling, C. L., K. L. Norman, and P. Sarnow.2006. Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122. Cold Spring Harb Sym 71:369-376.
8. Carone, D. M., M. S. Longo, G. C. Ferreri, L. Hall, M. Harris, N. Shook, K. V. Bulazel, B. R. Carone, C. Obergfell, M. J. O'Neill, and R. J. O'Neill.2009. A new class of retroviral and satellite encoded small RNAs emanates from mammalian centromeres. Chromosoma 118:113-125.
9. Cao, F., X. Z. Li, S. Hiew, H. Brady, Y. F. Liu, and Y. L. Dou.2009. Dicer independent small RNAs associate with telomeric heterochromatin. Rna 15:1274-1281.
10. Thompson, D. M., and R. Parker.2009. Stressing Out over tRNA Cleavage. Cell 138:215-219.
11. Taft, R. J., K. C. Pang, T. R. Mercer, M. Dinger, and J. S. Mattick.2010. Non-coding RNAs:regulators of disease. J Pathol 220:126-139.
12. Boehme, K. W., and T. Compton.2004. Innate sensing of viruses by toll-like receptors. J Virol 78:7867-7873.
13. Wieland, S. F., L. G. Guidotti, and F. V. Chisari.2000. Intrahepatic induction of alpha/beta interferon eliminates viral RNA-containing capsids in hepatitis B virus transgenic mice. J Virol 74:4165-4173.
14. Kimura, K., K. Kakimi, S. Wieland, L. G. Guidotti, and F. V. Chisari.2002. Activated intrahepatic antigen-presenting cells inhibit hepatitis B virus replication in the liver of transgenic mice. J Immunol 169:5188-5195.
15. Tang, T. J., J. Kwekkeboom, J. D. Laman, H. G. Niesters, P. E. Zondervan, R. A. de Man, S. W. Schalm, and H. L. Janssen.2003. The role of intrahepatic immune effector cells in inflammatory liver injury and viral control during chronic hepatitis B infection. Journal of viral hepatitis 10:159-167.
16. Kakimi, K., L. G. Guidotti, Y. Koezuka, and F. V. Chisari.2000. Natural killer T cell activation inhibits hepatitis B virus replication in vivo. Journal of Experimental Medicine 192:921-930.
17. Kimura, K., K. Kakimi, S. Wieland, L. G. Guidotti, and F. V. Chisari.2002. Interleukin-18 inhibits hepatitis B virus replication in the livers of transgenic mice. J Virol 76:10702-10707.
18. Pasquetto, V, L. G. Guidotti, K. Kakimi, M. Tsuji, and F. V. Chisari.2000. Host-virus interactions during malaria infection in hepatitis B virus transgenic mice. Journal of Experimental Medicine 192:529-535.
19. Wieland, S., R. Thimme, R. H. Purcell, and F. V. Chisari.2004. Genomic analysis of the host response to hepatitis B virus infection. Proc Natl Acad Sci U S A 101:6669-6674.
20. Wieland, S. F, and F. V. Chisari.2005. Stealth and cunning:Hepatitis B and hepatitis C viruses. J Virol 79:9369-9380.
21. Ganem, D.2004. Hepatitis B virus infection-Natural history and clinical consequences (vol 350, pg 1118,2004). New Engl J Med 351:1268-1268.
22. Summers, J.1988. The Replication Cycle of Hepatitis-B Viruses. Cancer 61:1957-1962.
23. Seeger, C, and W. S. Mason.2000. Hepatitis B virus biology. Microbiol Mol Biol R 64:51-+.
24. Ferrari, C, A. Bertoletti, A. Penna, A. Cavalli. A. Valli, G. Missale, M. Pilli, P. Fowler. T. Giuberti. F. V. Chisari. and F. Fiaccadori.1991. Identification of Immunodominant T-Cell Fpitopes of the Flepatitis-B Virus Nucleocapsid Antigen. Journal of Clinical Investigation 88:214-222.
25. Penna. A., M. Artini, A. Cavalli, M. Levrcro. A. Bertoletti. N. Pilli, F. V. Chisari. B. Rehermann. G. DelPrete. F. Fiaccadori, and C. Ferrari.1996. Long-lasting memory T cell responses following self-limited acute hepatitis B. Journal of Clinical Investigation 98:1185-1194.
26. Ferrari, C., A. Penna, A. Bertoletti, A. Valli, A. D. Antoni, T. Giuberti, A. Cavalli, M. A. Petit, and F. Fiaccadori.1990. Cellular Immune-Response to Hepatitis-B Virus-Encoded Antigens in Acute and Chronic Hepatitis-B Virus-Infection. J Immunol 145:3442-3449.
27. Mizukoshi, E., J. Sidney, B. Livingston, M. Ghany, J. H. Hoofnagle, A. Sette, and B. Rehermann.2004. Cellular immune responses to the hepatitis B virus polymerase. J Immunol 173:5863-5871.
28. Thimme, R., S. Wieland, C. Steiger, J. Ghrayeb, K. A. Reimann, R. H. Purcell, and F. V. Chisari.2003. CD8(+) T cells mediate viral clearance and disease pathogenesis during acute hepatitis B virus infection. J Virol 77:68-76.
29. Webster, G. J. M., S. Reignat, M. K. Maini, S. A. Whalley, G. S. Ogg, A. King, D. Brown, P. L. Amlot, R. Williams, D. Vergani, G. M. Dusheiko, and A. Bertoletti.2000. Incubation phase of acute hepatitis B in man:Dynamic of cellular immune mechanisms. Hepatology 32:1117-1124.
30. Biermer, M., R. Puro, and R. J. Schneider.2003. Tumor necrosis factor alpha inhibition of hepatitis B virus replication involves disruption of capsid integrity through activation of NF-kappa B. J Virol 77:4033-4042.
31. Guidotti, L. G., A. Morris, H. Mendez, R. Koch, R. H. Silverman, B. R. G. Williams, and F. V. Chisari.2002. Interferon-regulated pathways that control hepatitis B virus replication in transgenic mice. J Virol 76:2617-2621.
32. Guidotti, L. G., T. Ishikawa, M. V. Hobbs, B. Matzke, R. Schreiber, and F. V. Chisari. 1996. Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. Immunity 4:25-36.
33. Murray, J. M., S. F. Wieland, R. H. Purcell, and F. V. Chisari.2005. Dynamics of hepatitis B virus clearance in chimpanzees. Proc Natl Acad Sci U S A 102:17780-17785.
34. Maini, M. K., S. Reignat, C. Boni, G. S. Ogg, A. S. King, F. Malacarne, G. J. M. Webster, and A. Bertoletti.2000. T cell receptor usage of virus-specific CD8 cells and recognition of viral mutations during acute and persistent hepatitis B virus infection. Eur J Immunol 30:3067-3078.
35. Maini, M. K., C. Boni, C. K. Lee, J. R. Larrubia, S. Reignat, G. S. Ogg, A. S. King, J. Herberg, R. Gilson, A. Alisa, R. Williams, D. Vergani, N. V. Naoumov, C. Ferrari, and A. Bertoletti.2000. The role of virus-specific CD8(+) cells in liver damage and viral control during persistent hepatitis B virus infection. Journal of Experimental Medicine 191:1269-1280.
36. Sprengers, D., R. G. van der Molen,J. G. Kusters, R. A. De Man, H. G. M. Niesters, S. W. Schalm, and H. L. A. Janssen.2006. Analysis of intrahepatic HBV-specific cytotoxic T-cells during and after acute HBV infection in humans. J Hepatol 45:182-189.
37. Chisari, F. V., and C. Ferrari.1995. Hepatitis-B Virus Immunopathogenesis. Annual Review of Immunology 13:29-60.
38. Tang, T. J., J. Kwekkeboom, J. D. Laman, H. G. M. Niesters, P. E. Zondervan, R. A. de Man, S. W. Schalm, and H. L. A. Janssen.2003. The role of intrahepatic immune effector cells in inflammatory liver injury and viral control during chronic hepatitis B infection. Journal of viral hepatitis 10:159-167.
39. Chang, J. J., F. Wightman, A. Bartholomeusz, A. Ayres, S. J. Kent, J. Sasadeusz, and S. R. Lewin.2005. Reduced hepatitis B virus (HBV)-specific CD4+T-cell responses in human immunodeficiency virus type 1-HBV-coinfected individuals receiving HBV-active antiretroviral therapy. J Virol 79:3038-3051.
40. Reignat, S., G. J. Webster, D. Brown, G. S. Ogg, A. King, S. L. Seneviratne, G. Dusheiko, R. Williams, M. K. Maini, and A. Bertoletti.2002. Escaping high viral load exhaustion: CD8 cells with altered tetramer binding in chronic hepatitis B virus infection. J Exp Med 195:1089-1101.
41. Sprengers, D., R. G. van der Molen, J. G. Kusters, B. Hansen, H. G. Niesters, S. W. Schalm, and H. L. Janssen.2006. Different composition of intrahepatic lymphocytes in the immune-tolerance and immune-clearance phase of chronic hepatitis B. J Med Virol 78:561-568.
42. Boni, C., A. Penna, G. S. Ogg, A. Bertoletti, M. Pilli, C. Cavallo, A. Cavalli, S. Urbani, R. Boehme, R. Panebianco, F. Fiaccadori. and C. Ferrari.2001. Lamivudine treatment can overcome cytotoxic T-cell hyporesponsiveness in chronic hepatitis B:New perspectives for immune therapy. Hepatology 33:963-971.
43. Beckebaum, S., V. R. Cicinnati, X. Zhang, S. Ferencik, A. Frilling, H. Grosse-Wilde, C. E. Broelsch, and G. Gerken.2003. Hepatitis B virus-induced defect of monocyte-derived dendritic cells leads to impaired T helper type 1 response in vitro:mechanisms for viral immune escape. Immunology 109:487-495.
44. Trautmann. L.. L. Janbazian. N. Chomont, E. A. Said. S. Gimmig. B. Bessette. M. R. Boulassel. E. Delwarl, H. Sepulveda, R. S. Balderas. J. P. Routy. E. K. Haddad. and R. P. Sekaly.2006. Upregulation of PD-1 expression on ⅢⅤ-specific CD8(+) T cells leads to reversible immune dysfunction (vol 12.1198.2006). Nature Medicine 12:1329-1329.
45. Barber, D. L., E. J. Wherry, D. Masopust, B. G. Zhu, J. P. Allison, A. H. Sharpe, G. J. Freeman, and R. Ahmed.2006. Restoring function in exhausted CD8 T cells during chronic viral infection. Nature 439:682-687.
46. Boettler, T., E. Panther, B. Bengsch, N. Nazarova, H. C. Spangenberg, H. E. Blum, and R. Thimme.2006. Expression of the interleukin-7 receptor alpha chain (CD 127) on virus-specific CD8(+) T cells identifies functionally and phenotypically defined memory T cells during acute resolving hepatitis B virus infection. J Virol 80:3532-3540.
47. Akdis, M., K. Blaser, and C. A. Akdis.2005. T regulatory cells in allergy:novel concepts in the pathogenesis, prevention, and treatment of allergic diseases. J Allergy Clin Immunol 116:961-968; quiz 969.
48. Franzese, O., P. T. Kennedy, A. J. Gehring, J. Gotto, R. Williams, M. K. Maini, and A. Bertoletti.2005. Modulation of the CD8+-T-cell response by CD4+CD25+ regulatory T cells in patients with hepatitis B virus infection. J Virol 79:3322-3328.
49. Stoop, J. N., R. G. van der Molen, C. C. Baan, L. J. W. van der Laan, E. J. Kuipers, J. G. Kusters, and H. L. A. Janssen.2005. Regulatory T cells contribute to the impaired immune response in patients with chronic hepatitis B virus infection. Hepatology 41:771-778.
50. Beckebaum, S., V. R. Cicinnati, G. Dworacki, J. Muller-Berghaus, D. Stolz, J. Harnaha, T. L. Whiteside, A. W. Thomson, L. Lu, J. J. Fung, and C. A. Bonham.2002. Reduction in the circulating pDC1/pDC2 ratio and impaired function of ex vivo-generated DC1 in chronic hepatitis B infection. Clin Immunol 104:138-150.
51. Zheng, B. J., J. Zhou, D. Qu, K. L. Siu, T. W. Lam, H. Y. Lo, S. S. Lee, and Y. M. Wen. 2004. Selective functional deficit in dendritic cell-T cell interaction is a crucial mechanism in chronic hepatitis B virus infection. Journal of viral hepatitis 11:217-224.
52. van der Molen, R. G., D. Sprengers, R. S. Binda, E. C. de Jong, H. G. M. Niesters, J. G. Kusters, J. Kwekkeboom, and H. L. A. Janssen.2004. Functional impairment of myeloid and plasmacytoid dendritic cells of patients with chronic hepatitis B. Hepatology 40:738-746.
53. Riordan, S. M., N. Skinner, J. Kurtovic, S. Locarnini, and K. Visvanathan.2006. Reduced expression of toll-like receptor 2 on peripheral monocytes in patients with chronic hepatitis B. Clinical and Vaccine Immunology 13:972-974.
54. Ribeiro, R. M., J. Layden-Almer, K. A. Powers, T. J. Layden, and A. S. Perelson.2003. Dynamics of alanine aminotransferase during hepatitis C virus treatment. Hepatology 38:509-517.
55. Tedder, R. S., S. Ijaz, N. Gilbert, J. A. Barbara, S. A. Corden, R. J. Gilson, and E. H. Boxall.2002. Evidence for a dynamic host-parasite relationship in e-negative hepatitis B carriers. J Med Virol 68:505-512.
56. Liaw, Y. F., R. N. Chien, C. T. Yeh, S. L. Tsai, and C. M. Chu.1999. Acute exacerbation and hepatitis B virus clearance after emergence of YMDD motif mutation during lamivudine therapy. Hepatology 30:567-572.
57. Milich, D., and T. J. Liang.2003. Exploring the biological basis of hepatitis B e antigen in hepatitis B virus infection. Hepatology 38:1075-1086.
58. Chen, M., M. Sallberg, S. N. Thung, J. Hughes, J. Jones, and D. R. Milich.2000. Nondeletional T-cell receptor transgenic mice:model for the CD4(+) T-cell repertoire in chronic hepatitis B virus infection. J Virol 74:7587-7599.
59. Rossol, S., G. Marinos, P. Carucci, M. V. Singer, R. Williams, and N. V. Naoumov.1997. Interleukin-12 induction of Thl cytokines is important for viral clearance in chronic hepatitis B. Journal of Clinical Investigation 99:3025-3033.
60. Tang, T. J., H. L. A. Janssen, J. G. Kusters, R. A. de Man, S. W. Schalm, and J. Kwekkeboom.2003. The intrahepatic immune response during chronic hepatitis B infection can be monitored by the fine-needle aspiration biopsy technique. Fems Immunology and Medical Microbiology 39:69-72.
61. Bonecchi, R., G. Bianchi, P. P. Bordignon, D. D'Ambrosio, R. Lang, A. Borsatti, S. Sozzani, P. Allavena, P. A. Gray, A. Mantovani, and F. Sinigaglia.1998. Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Thls) and Th2s. J Exp Med 187:129-134.
62. Friedman, S. L.2000. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. Journal of Biological Chemistry 275:2247-2250.
63. Canbay, A., P. Taimr, N. Torok, H. Higuchi, S. Friedman, and G. J. Gores.2003. Apoptotic body engulfment by a human stellate cell line is profibrogenic. Lab Invest 83:655-663.
64. Jo. M., T. H. Kim, D. W. Seol, J. E. Esplen. K. Dorko. T. R. Billiar, and S. C. Strom.2000. Apoptosis induced in normal human hepatocytes by tumor necrosis factor-related apoptosis-inducing ligand. Nature Medicine 6:564-567.
65. Su. F. and R.J. Schneider.1997. Hepatitis B virus HBx protein sensitizes cells to apopiotic killing by tumor necrosis factor alpha. Proc Natl Acad Sci U S A 94:8744-8749.
66. Janssen, H. L. A.,H. Higuchi,A. Abdulkarim, and G. J. Gores.2003. Hepatitis B virus enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) cytotoxicity by increasing TRAIL-R1/death receptor 4 expression. J Hepatol 39:414-420.
67. Lee, J. Y., D. W. Chae, S. M. Kim, E. S. Nam, M. K. Jang, J. H. Lee, H. Y. Kim, and J. Y. Yoo.2004. Expression of FasL and perforin/granzyme B mRNA in chronic hepatitis B virus infection. Journal of viral hepatitis 11:130-135.
68. Luo, K. X., Y. F. Zhu, L. X. Zhang, H. T. He, X. S. Wang, and L. Zhang. 1997.In situ investigation of Fas/FasL expression in chronic hepatitis B infection and related liver diseases. Journal of viral hepatitis 4:303-307.
69. Rivero, M., J. Crespo, E. Fabrega, F. Casafont, M. Mayorga, M. Gomez-Fleitas, and F. Pons-Romero.2002. Apoptosis mediated by the Fas system in the fulminant hepatitis by hepatitis B virus. Journal of viral hepatitis 9:107-113.
70. Lai, C. L., and M. F. Yuen.2013. Prevention of hepatitis B virus-related hepatocellular carcinoma with antiviral therapy. Hepatology 57:399-408.
71. Chisari, F. V.2000. Rous-Whipple Award Lecture. Viruses, immunity, and cancer:lessons from hepatitis B. Am J Pathol 156:1117-1132.
72. Guidotti, L. G., and F. V. Chisari.2006. Immunobiology and pathogenesis of viral hepatitis. Annu Rev Pathol 1:23-61.
73. Kremsdorf, D., P. Soussan, P. Paterlini-Brechot, and C. Brechot.2006. Hepatitis B virus-related hepatocellular carcinoma:paradigms for viral-related human carcinogenesis. Oncogene 25:3823-3833.
74. Matsubara, K., and T. Tokino.1990. Integration of hepatitis B virus DNA and its implications for hepatocarcinogenesis. Mol Biol Med 7:243-260.
75. Minami, M., Y. Daimon, K. Mori, H. Takashima, T. Nakajima, Y. Itoh, and T. Okanoue. 2005. Hepatitis B virus-related insertional mutagenesis in chronic hepatitis B patients as an early drastic genetic change leading to hepatocarcinogenesis. Oncogene 24:4340-4348.
76. Kekule, A. S., U. Lauer, L. Weiss, B. Luber, and P. H. Hofschneider.1993. Hepatitis B virus transactivator HBx uses a tumour promoter signalling pathway. Nature 361:742-745.
77. Maguire, H. F., J. P. Hoeffler, and A. Siddiqui.1991. HBV X protein alters the DNA binding specificity of CREB and ATF-2 by protein-protein interactions. Science 252:842-844.
78. Natoli, G., M. L. Avantaggiati, P. Chirillo, A. Costanzo, M. Artini, C. Balsano, and M. Levrero.1994. Induction of the DNA-binding activity of c-jun/c-fos heterodimers by the hepatitis B virus transactivator pX. Mol Cell Biol 14:989-998.
79. Brechot, C.2004. Pathogenesis of hepatitis B virus-related hepatocellular carcinoma:old and new paradigms. Gastroenterology 127:S56-61.
80. lavarone, M., J. B. Trabut, O. Delpuech, F. Carnot, M. Colombo, D. Kremsdorf. C. Brechot, and V. Thiers.2003. Characterisation of hepatitis B virus X protein mutants in tumour and non-tumour liver cells using laser capture microdissection. J Hepatol 39:253-261.
81. Leon, B., L. Taylor, M. Vargas, R. B. Luftig, F. Albertazzi, L. Herrero, and K. Visona. 2005. HBx M130K and V1311 (T-A) mutations in HBV genotype F during a follow-up study in chronic carriers. Virol J 2:60.
82. Caselmann, W. H., M. Meyer, A. S. Kekule, U. Lauer, P. H. Hofschneider, and R. Koshy. 1990. A trans-activator function is generated by integration of hepatitis B virus preS/S sequences in human hepatocellular carcinoma DNA. Proc Natl Acad Sci U S A 87:2970-2974.
83. Kekule, A. S., U. Lauer, M. Meyer, W. H. Caselmann, P. H. Hofschneider, and R. Koshy. 1990. The preS2/S region of integrated hepatitis B virus DNA encodes a transcriptional transactivator. Nature 343:457-461.
84. Hildt, E., G. Saher, V. Bruss, and P. H. Hofschneider.1996. The hepatitis B virus large surface protein (LHBs) is a transcriptional activator. Virology 225:235-239.
85. Hagen, T. M., S. Huang, J. Curnutte, P. Fowler, V. Martinez, C. M. Wehr, B. N. Ames, and F. V. Chisari.1994. Extensive oxidative DNA damage in hepatocytes of transgenic mice with chronic active hepatitis destined to develop hepatocellular carcinoma. Proc Natl Acad Sci U S A 91:12808-12812.
86. Huang, S. N., and F. V. Chisari.1995. Strong, sustained hepatocellular proliferation precedes hepatocarcinogenesis in hepatitis B surface antigen transgenic mice. Hepatology 21:620-626.
87. Nakamoto, Y., L. G. Guidotti, C. V. Kuhlen, P. Fowler, and F. V. Chisari.1998. Immune pathogenesis of hepatocellular carcinoma. J Exp Med 188:341-350.
88. Baumert, T. E, R. Thimme, and F. von Weizsacker.2007. Pathogenesis of hepatitis B virus infection. World J Gastroenterol 13:82-90.
89. Liang, T. J., K. Hasegawa. N. Rimon, J. R. Wands, and E. Ben-Porath.1991. A hepatitis B virus mutant associated with an epidemic of fulminant hepatitis. N Engl J Med 324:1705-1709.
90. Baumert. T. F.. S. A. Rogers. K. Hasegawa, and T. J. Liang.1996. Two core promotor mutations identified in a hepatitis B virus strain associated with fulminant hepatitis result in enhanced viral replication..1 Clin Invest 98:2268-2276.
91. Baumert, T. F., C. Yang, P. Schurmann, J. Kock, C. Ziegler, C. Grullich, M. Nassal, T. J. Liang, H. E. Blum, and F. von Weizsacker.2005. Hepatitis B virus mutations associated with fulminant hepatitis induce apoptosis in primary Tupaia hepatocytes. Hepatology 41:247-256.
92. Buckwold, V. E., Z. Xu, M. Chen, T. S. Yen,and J. H. Ou. 1996.Effects of a naturally occurring mutation in the hepatitis B virus basal core promoter on precore gene expression and viral replication. J Virol 70:5845-5851.
93. Zoulim, F.2004. Mechanism of viral persistence and resistance to nucleoside and nucleotide analogs in chronic hepatitis B virus infection. Antiviral Res 64:1-15.
94. Schildgen, O., H. Sirma, A. Funk, C. Olotu, U. C. Wend, H. Hartmann, M. Helm, J. K. Rockstroh, W. R. Willems, H. Will, and W. H. Gerlich.2006. Variant of hepatitis B virus with primary resistance to adefovir. N Engl J Med 354:1807-1812.
95. Baumert, T. F., H. Barth, and H. E. Blum.2005. Genetic variants of hepatitis B virus and their clinical relevance. Minerva Gastroenterol Dietol 51:95-108.
96. Gouas, D. A., S. Villar, S. Ortiz-Cuaran, P. Legros, G. Ferro, G. D. Kirk, O. A. Lesi, M. Mendy, E. Bah, M. D. Friesen, J. Groopman, I. Chemin, and P. Hainaut.2012. TP53 R249S mutation, genetic variations in HBX and risk of hepatocellular carcinoma in The Gambia. Carcinogenesis 33:1219-1224.
97. Ma, N. F., S. H. Lau, L. Hu, D. Xie, J. Wu, J. Yang, Y. Wang, M. C. Wu, J. Fung, X. Bai, C. H. Tzang, L. Fu, M. Yang, Y. A. Su, and X. Y. Guan.2008. COOH-terminal truncated HBV X protein plays key role in hepatocarcinogenesis. Clin Cancer Res 14:5061-5068.
98. Xu, R., X. Zhang, W. Zhang, Y. Fang, S. Zheng, and X. F. Yu.2007. Association of human APOBEC3 cytidine deaminases with the generation of hepatitis virus B x antigen mutants and hepatocellular carcinoma. Hepatology 46:1810-1820.
99. Carey, I., L. D'Antiga, S. Bansal, M. S. Longhi, Y. Ma, I. R. Mesa, G. Mieli-Vergani, and D. Vergani.2011. Immune and viral profile from tolerance to hepatitis B surface antigen clearance:a longitudinal study of vertically hepatitis B virus-infected children on combined therapy. J Virol 85:2416-2428.
100. Brunetto, M. R., M. Stemler, F. Bonino, F. Schodel, F. Oliveri, M. Rizzetto, G. Verme, and H. Will.1990. A new hepatitis B virus strain in patients with severe anti-HBe positive chronic hepatitis B. J Hepatol 10:258-261.
101. Akahane, Y., T. Yamanaka, H. Suzuki, Y. Sugai, F. Tsuda, S. Yotsumoto, S. Omi, H. Okamoto, Y. Miyakawa, and M. Mayumi.1990. Chronic Active Hepatitis with Hepatitis-B Virus-DNA and Antibody against E-Antigen in the Serum-Disturbed Synthesis and Secretion of E-Antigen from Hepatocytes Due to a Point Mutation in the Precore Region. Gastroenterology 99:1113-1119.
102. Milich, D. R.1999. Do T cells "see" the hepatitis B core and e antigens differently? Gastroenterology 116:765-768.
103. Diepolder, H. M., G. Ries, M. C. Jung, H. J. Schlicht, J. T. Gerlach, N. Gr ner, W. H. Caselmann, and G. R. Pape.1999. Differential antigen-processing pathways of the hepatitis B virus e and core proteins. Gastroenterology 116:650-657.
104. Tacke, F., C. Gehrke, T. Luedde, A. Heim, M. P. Manns, and C. Trautwein.2004. Basal core promoter and precore mutations in the hepatitis B virus genome enhance replication efficacy of Lamivudine-resistant mutants. J Virol 78:8524-8535.
105. Parekh, S., F. Zoulim, S. H. Ahn, A. Tsai, J. Li, S. Kawai, N. Khan, C. Trepo, J. Wands, and S. Tong.2003. Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants. J Virol 77:6601-6612.
106. Preikschat, P., S. Gunther, S. Reinhold, H. Will, K. Budde, H. H. Neumayer, D. H. Kruger, and H. Meisel.2002. Complex HBV populations with mutations in core promoter, C gene, and pre-S region are associated with development of cirrhosis in long-term renal transplant recipients. Hepatology 35:466-477.
107. Sato, S., K. Suzuki, Y. Akahane, K. Akamatsu, K. Akiyama, K. Yunomura, F. Tsuda, T. Tanaka, H. Okamoto, Y. Miyakawa, and M. Mayumi.1995. Hepatitis B virus strains with mutations in the core promoter in patients with fulminant hepatitis. Ann Intern Med 122:241-248.
108. Okamoto, H., F. Tsuda, Y. Akahane, Y. Sugai, M. Yoshiba, K. Moriyama, T. Tanaka, Y. Miyakawa, and M. Mayumi.1994. Hepatitis B virus with mutations in the core promoter for an e antigen-negative phenotype in carriers with antibody to e antigen. J Virol 68:8102-8110.
109. Li, J., V. E. Buckwold, M. W. Hon, and J. H. Ou.1999. Mechanism of suppression of hepatitis B virus precore RNA transcription by a frequent double mutation. J Virol 73:1239-1244.
110. Gunther, S., N. Piwon, A. Iwanska, R. Schilling. H. Meisel, and H. Will.1996. Type, prevalence, and significance of core promoter/enhancer Ⅱ mutations in hepatitis B viruses from immunosuppressed patients with severe liver disease. J Virol 70:8318-8331.
111. Pult,L. T. Chouard. S. Wieland, R. Klemenz. M. Yaniv, and H. E. Blum.1997. A hepatitis B virus mutant with a new hepatocyte nuclear factor 1 binding site emerging in transplant-transmitted fulminant hepatitis B. Hepatology 25:1507-1515.
112. Zheng, Y., J. Li, and J. H. Ou.2004. Regulation of hepatitis B virus core promoter by transcription factors HNF1 and HNF4 and the viral X protein. J Virol 78:6908-6914.
113. Kao, J. H., P. J. Chen, M. Y. Lai, and D. S. Chen.2003. Basal core promoter mutations of hepatitis B virus increase the risk of hepatocellular carcinoma in hepatitis B carriers. Gastroenterology 124:327-334.
114. Baumert, T. F., A. Marrone, J. Vergalla, and T. J. Liang.1998. Naturally occurring mutations define a novel function of the hepatitis B virus core promoter in core protein expression. J Virol 72:6785-6795.
115. Zuckerman, J. N., and A. J. Zuckerman.2003. Mutations of the surface protein of hepatitis B virus. Antiviral Res 60:75-78.
116. Carman, W. F., A. R. Zanetti, P. Karayiannis, J. Waters, G. Manzillo, E. Tanzi, A. J. Zuckerman, and H. C. Thomas.1990. Vaccine-induced escape mutant of hepatitis B virus. Lancet 336:325-329.
117. Waters, J. A., M. Kennedy, P. Voet, P. Hauser, J. Petre, W. Carman, and H. C. Thomas. 1992. Loss of the common "A" determinant of hepatitis B surface antigen by a vaccine-induced escape mutant. J Clin Invest 90:2543-2547.
118. Kalinina, T., A. Iwanski, H. Will, and M. Sterneck.2003. Deficiency in virion secretion and decreased stability of the hepatitis B virus immune escape mutant G145R. Hepatology 38:1274-1281.
119. Ogata, N., P. J. Cote, A. R. Zanetti, R. H. Miller, M. Shapiro, J. Gerin, and R. H. Purcell. 1999. Licensed recombinant hepatitis B vaccines protect chimpanzees against infection with the prototype surface gene mutant of hepatitis B virus. Hepatology 30:779-786.
120. Bauer, T, K. Weinberger, and W. Jilg.2002. Variants of two major T cell epitopes within the hepatitis B surface antigen are not recognized by specific T helper cells of vaccinated individuals. Hepatology 35:455-465.
121. Protzer-Knolle, U., U. Naumann, R. Bartenschlager, T. Berg, U. Hopf, K. H. M. zum Buschenfelde, P. Neuhaus, and G. Gerken.1998. Hepatitis B virus with antigenically altered hepatitis B surface antigen is selected by high-dose hepatitis B immune globulin after liver transplantation. Hepatology 27:254-263.
122. Ghany, M. G., B. Ayola, F. G. Villamil, R. G. Gish, S. Rojter, J. M. Vierling, and A. S. F. Lok.1998. Hepatitis B virus S mutants in liver transplant recipients who were reinfected despite hepatitis B immune globulin prophylaxis. Hepatology 27:213-222.
123. Bes, M., V. Vargas, M. Piron, N. Casamitjana, J. I. Esteban, N. Vilanova, A. Pinacho, J. Quer, L. Puig, J. Guardia, and S. Sauleda.2012. T cell responses and viral variability in blood donation candidates with occult hepatitis B infection. J Hepatol 56:765-774.
124. Pollicino, T., G. Raffa, L. Costantino, A. Lisa, C. Campello, G. Squadrito, M. Levrero, and G. Raimondo.2007. Molecular and functional analysis of occult hepatitis B virus isolates from patients with hepatocellular carcinoma. Hepatology 45:277-285.
125. El Chaar, M., D. Candotti, R. A. Crowther, and J. P. Allain.2010. Impact of Hepatitis B Virus Surface Protein Mutations on the Diagnosis of Occult Hepatitis B Virus Infection. Hepatology 52:1600-1610.
126. Yuen, M. F., E. Sablon, C. K. Hui, H. J. Yuan, H. Decraemer, and C. L. Lai.2001. Factors associated with hepatitis B virus DNA breakthrough in patients receiving prolonged lamivudine therapy. Hepatology 34:785-791.
127. Lai, C. L., J. Dienstag, E. Schiff, N. W. Leung, M. Atkins, C. Hunt, N. Brown, M. Woessner, R. Boehme, and L. Condreay.2003. Prevalence and clinical correlates of YMDD variants during lamivudine therapy for patients with chronic hepatitis B. Clin Infect Dis 36:687-696.
128. Marcellin, P., T. T. Chang, S. G. Lim, M. J. Tong, W. Sievert, M. L. Shiffman, L. Jeffers, Z. Goodman, M. S. Wulfsohn, S. Xiong, J. Fry, and C. L. Brosgart.2003. Adefovir dipivoxil for the treatment of hepatitis B e antigen-positive chronic hepatitis B. N Engl J Med 348:808-816.
129. Hadziyannis, S. J., N. C. Tassopoulos, E. J. Heathcote, T. T. Chang, G. Kitis, M. Rizzetto, P. Marcellin, S. G. Lim, Z. Goodman, M. S. Wulfsohn, S. Xiong, J. Fry, and C. L. Brosgart.2003. Adefovir dipivoxil for the treatment of hepatitis B e antigen-negative chronic hepatitis B. N Engl J Med 348:800-807.
130. Liaw, Y. F., J.J. Sung, W. C. Chow, G. Farrell, C. Z. Lee, H. Yuen, T. Tanwandee, Q. M. Tao, K. Shue, O. N. Keene, J. S. Dixon, D. F. Gray, and J. Sabbat.2004. Lamivudine for patients with chronic hepatitis B and advanced liver disease. N Engl J Med 351:1521-1531.
131. Lok, A. S., C. L. Lai, N. Leung, G. B. Yao, Z. Y. Cui, E. R. Schiff. J. L. Dienstag, E. J. Heathcote, N. R. Little, D. A. Griffiths, S. D. Gardner, and M. Castiglia.2003. Long-term safety of lamivudine treatment in patients with chronic hepatitis B. Gastroenterology 125:1714-1722.
132. Allen, M. I., M. Deslauriers. C. W. Andrews. G. A. Tipples, K. A. Walters. D. L. Tyrrell, N. Brown, and L. D. Condreay.1998. Identification and characterization of mutations in hepatitis B virus resistant to lamivudine. Lamivudine Clinical Investigation Group. Hepatology 27:1670-1677.
133. Melegari, M., P. P. Scaglioni, and J. R. Wands.1998. Hepatitis B virus mutants associated with 3TC and famciclovir administration are replication defective. Hepatology 27:628-633.
134. Delaney, W. E. t., H. Yang, C. E. Westland, K. Das, E. Arnold, C. S. Gibbs, M. D. Miller, and S. Xiong.2003. The hepatitis B virus polymerase mutation rtV173L is selected-during lamivudine therapy and enhances viral replication in vitro. J Virol 77:11833-11841.
135. Benhenda, S., D. Cougot, M. A. Buendia, and C. Neuveut.2009. Hepatitis B virus X protein molecular functions and its role in virus life cycle and pathogenesis. Adv Cancer Res 103:75-109.
136. Feitelson, M. A., and J. Lee.2007. Hepatitis B virus integration, fragile sites, and hepatocarcinogenesis. Cancer Lett 252:157-170.
137. Jiang, W., X. W. Wang, T. Unger, M. Forgues, J. W. Kim, S. P. Hussain, E. Bowman, E. A. Spillare, M. M. Lipsky, J. M. Meck, L. R. Cavalli, B. R. Haddad, and C. C. Harris.2010. Cooperation of tumor-derived HBx mutants and p53-249(ser) mutant in regulating cell proliferation, anchorage-independent growth and aneuploidy in a telomerase-immortalized normal human hepatocyte-derived cell line. Int J Cancer 127:1011-1020.
138. Boni, C., A. Penna, G. S. Ogg, A. Bertoletti, M. Pilli, C. Cavallo, A. Cavalli, S. Urbani, R. Boehme, R. Panebianco, F. Fiaccadori, and C. Ferrari.2001. Lamivudine treatment can overcome cytotoxic T-cell hyporesponsiveness in chronic hepatitis B:new perspectives for immune therapy. Hepatology 33:963-971.
139. Kim, D., K. S. Lyoo, D. Smith, W. Hur, S. W. Hong, P. S. Sung, S. K. Yoon, and S. Mehta. 2011. Number of Mutations Within CTL-Defined Epitopes of the Hepatitis B Virus (HBV) Core Region is Associated With HBV Disease Progression. J Med Virol 83:2082-2087.
140. Simmonds, P., and S. Midgley.2005. Recombination in the genesis and evolution of hepatitis B virus genotypes. J Virol 79:15467-15476.
141. Sobao, Y., K. Sugi, H. Tomiyama, S. Saito, S. Fujiyama, M. Morimoto, S. Hasuike, H. Tsubouchi, K. Tanaka, and M. Takiguch.2001. Identification of hepatitis B virus-specific CTL epitopes presented by HLA-A*2402, the most common HLA class I allele in East Asia. J Hepatol 34:922-929.
142. Chisari, F. V, and C. Ferrari.1995. Hepatitis B virus immunopathogenesis. Annu Rev Immunol 13:29-60.
143. Chang, J. J., and S. R. Lewin.2007. Immunopathogenesis of hepatitis B virus infection. Immunol Cell Biol 85:16-23.
144. Parker, D. C.1993. T cell-dependent B cell activation. Annu Rev Immunol 11:331-360.
145. Yewdell, J. W., and J. R. Bennink.1992. Cell biology of antigen processing and presentation to major histocompatibility complex class I molecule-restricted T lymphocytes. Adv Immunol 52:1-123.
146. Vitetta, E. S., R. Fernandez-Botran, C. D. Myers, and V. M. Sanders.1989. Cellular interactions in the humoral immune response. Adv Immunol 45:1-105.
147. Doherty, P. C., W. Allan, M. Eichelberger, and S. R. Carding.1992. Roles of alpha beta and gamma delta T cell subsets in viral immunity. Annu Rev Immunol 10:123-151.
148. Bertoni, R., J. Sidney, P. Fowler, R. W. Chesnut, F. V. Chisari, and A. Sette.1997. Human histocompatibility leukocyte antigen-binding supermotifs predict broadly cross-reactive cytotoxic T lymphocyte responses in patients with acute hepatitis. J Clin Invest 100:503-513.
149. Lee, W. M.1997. Hepatitis B virus infection. N Engl J Med 337:1733-1745.
150. Hwang, Y. K., N. K. Kim, J. M. Park, K. Lee, W. K. Han, H. I. Kim, and H. S. Cheong. 2002. HLA-A21 restricted peptides from the HBx antigen induce specific CTL responses in vitro and in vivo. Vaccine 20:3770-3777.
151. Rehennann, B., P. Fowler, J. Sidney, J. Person, A. Redeker, M. Brown, B. Moss, A. Sette, and F. V. Chisari.1995. The cytotoxic T lymphocyte response to multiple hepatitis B virus polymerase epitopes during and after acute viral hepatitis. J Exp Med 181:1047-1058.
152. Mizukoshi, E., J. Sidney, B. Livingston, M. Ghany, J. H. Hoofnagle, A. Sette, and B. Rehermann.2004. Cellular immune responses to the hepatitis B virus polymerase. J Immunol 173:5863-5871.
153. Letsch, A., and C. Scheibenbogen.2003. Quantification and characterization of specific T-cells by antigen-specific cytokine production using ELISPOT assay or intracellular cytokine staining. Methods 31:143-149.
154. Ferrari, C., A. Bertoletti, A. Penna, A. Cavalli, A. Valli, G. Missale, M. Pilli, P. Fowler, T. Giuberti, F. V. Chisari, and et al.1991. Identification of immunodominant T cell epitopes of the hepatitis B virus nucleocapsid antigen. J Clin Invest 88:214-222.
155. Bertoletti, A., C. Ferrari, F. Fiaccadori. A. Penna, R. Margolskee. H.J. Schlicht. P. Fowler, S. Guilhot, and F. V. Chisari.1991. HLA class I-restricted human cytotoxic T cells recognize endogenously synthesized hepatitis B virus nucleocapsid antigen. Proc Natl Acad Sci U S A 88:10445-10449.
156. Penna. A., F. V. Chisari. A. Bertoletti, G. Missale, P. Fowler. T. Giuberti. F. Fiaccadori, and C. Ferrari.1991. Cytotoxic T lymphocytes recognize an HLA-A2-restricted epitope within the hepatitis B virus nucleocapsid antigen. J Exp Med 174:1565-1570.
157. Betts, M. R., D. R. Ambrozak, D. C. Douek, S. Bonhoeffer, J. M. Brenchley, J. P. Casazza, R. A. Koup, and L. J. Picker.2001. Analysis of total human immunodeficiency virus (HIV)-specific CD4(+) and CD8(+) T-cell responses:responship to viral load in untreated HIV infection. J Virol 75:11983-11991.
158. Rammensee, H. G., K. Falk, and O. Rotzschke.1993. Peptides naturally presented by MHC class I molecules. Annu Rev Immunol 11:213-244.
159. Falk, K., O. Rotzschke, S. Stevanovic, G. Jung, and H. G. Rammensee.1991. Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules. Nature 351:290-296.
160. Tsai, S. L., M. H. Chen, C. T. Yeh, C. M. Chu, A. N. Lin, F. H. Chiou, T. H. Chang, and Y. F. Liaw.1996. Purification and characterization of a naturally processed hepatitis B virus peptide recognized by CD8+ cytotoxic T lymphocytes. J Clin Invest 97:577-584.
161. Meng, S. D., T. Gao, G. F. Gao, and P. Tien.2001. HBV-specific peptide associated with heat-shock protein gp96. Lancet 357:528-529.
162. Li, H., M. Zhou, J. Han, X. Zhu, T. Dong, G. F. Gao, and P. Tien.2005. Generation of murine CTL by a hepatitis B virus-specific peptide and evaluation of the adjuvant effect of heat shock protein glycoprotein 96 and its terminal fragments. J Immunol 174:195-204.
163. Cao, T, I. Desombere, P. Vanlandschoot, M. Sallberg, and G. Leroux-Roels.2002. Characterization of HLA DR13-restricted CD4(+) T cell epitopes of hepatitis B core antigen associated with self-limited, acute hepatitis B. J Gen Virol 83:3023-3033.
164. Desombere, I., Y. Gijbels, A. Verwulgen, and G. Leroux-Roels.2000. Characterization of the T cell recognition of hepatitis B surface antigen (HBsAg) by good and poor responders to hepatitis B vaccines. Clin Exp Immunol 122:390-399.
165. Pajot, A., M. L. Michel, M. Mancini-Bourgine, M. N. Ungeheuer, D. M. Ojcius, Q. Deng, F. A. Lemonnier, and Y. C. Lone.2006. Identification of novel HLA-DR1-restricted epitopes from the hepatitis B virus envelope protein in mice expressing HLA-DR1 and vaccinated human subjects. Microbes Infect 8:2783-2790.
166. Doh, H., S. Roh, K. W. Lee, and K. Kim.2003. Response of primed human PBMC to synthetic peptides derived from hepatitis B virus envelope proteins:a search for promiscuous epitopes. FEMS Immunol Med Microbiol 35:77-85.
167. Honorati, M. C., P. Dolzani, E. Mariani, A. Piacentini, G. Lisignoli, C. Ferrari, and A. Facchini.1997. Epitope specificity of Th0/Th2 CD4+ T-lymphocyte clones induced by vaccination with rHBsAg vaccine. Gastroenterology 112:2017-2027.
168. Lin, C. L., S. L. Tsai, T. H. Lee, R. N. Chien, S. K. Liao, and Y. F. Liaw.2005. High frequency of functional anti-YMDD and -mutant cytotoxic T lymphocytes after in vitro expansion correlates with successful response to lamivudine therapy for chronic hepatitis B. Gut 54:152-161.
169. Lee, H. G., J. S. Lim, K. Y. Lee, Y. K. Choi, I. S. Choe, T. W. Chung, and K. Kim.1997. Peptide-specific CTL induction in HBV-seropositive PBMC by stimulation with peptides in vitro:novel epitopes identified from chronic carriers. Virus Res 50:185-194.
170. Bertoletti, A., S. Southwood, R. Chesnut, A. Sette, M. Falco, G. B. Ferrara, A. Penna, C. Boni, F. Fiaccadori, and C. Ferrari.1997. Molecular features of the hepatitis B virus nucleocapsid T-cell epitope 18-27:interaction with HLA and T-cell receptor. Hepatology 26:1027-1034.
171. Thimme, R., K. M. Chang, J. Pemberton, A. Sette, and F. V. Chisari.2001. Degenerate immunogenicity of an HLA-A2-restricted hepatitis B virus nucleocapsid cytotoxic T-lymphocyte epitope that is also presented by HLA-B51. J Virol 75:3984-3987.
172. Bertoletti, A., and A. J. Gehring.2006. The immune response during hepatitis B virus infection. J Gen Virol 87:1439-1449.
1 Thimme R, Wieland S, Steiger C, Ghrayeb J, Reimann KA, Purcell RH, Chisari FV.2003. CD8(+) T cells mediate viral clearance and disease pathogenesis during acute hepatitis B virus infection. J Virol.77:68-76.
2 Maini M, Boni C, Lee C, Larrubia J, Reignat S, Ogg G et al.2000. The role of virus-specific CD8(+) cells in liver damage and viral control during persistent hepatitis B virus infection. J Exp Med.191:1269.
3 Chang JJ, Wightman F, Bartholomeusz A, Ayres A, Kent SJ, Sasadeusz J et al.2005. Reduced HBV-specific CD4+T-cell responses in HIV-1-HBV co-infected individuals receiving HBV-active antiretroviral therapy. J Virol.79:3038-3051.
4 George J. M. Webster, Stephanie Reignat, David Brown et al.2004. Longitidinal 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.
5 Andersen M H, Schrama D, Thor Straten P, et al. Cytotoxic T cells (J).2006. J Invest Dermatol.126:32-41.
6 Rehermann, B., P. Fowler, J. Sidney, J. Person, A. Redeker, M. Brown, B. Moss, A. Sette, and F. V. Chisari.1995. The cytotoxic T lymphocyte response to multiple hepatitis B vims polymerase epitopes during and after acute viral hepatitis. J. Exp. Med.181:1047-1058.
7 Sobao, Y., K. Sugi, H. Tomiyama, S. Saito, S. Fujiyama, M. Morimoto, S. Hasuike, H. Tsubouchi, K. Tanaka, and M. Takiguchi.2001. Identification of hepatitis B virus-specific CTL epitopes presented by HLA-A*2402, the most common HLA class I allele in East Asia. J. Hepatol.34:922-929.
8 Nayersina, R., P. Fowler, S. Guilhot, G. Missale, A. Cerny, H. J. Schlicht, A. Vitiello, R. Chesnut, J. L. Person, A. J. Redeker, and F. V. Chisari.1993. HLA-A2 restricted cytotoxic T lymphocyte responses to multiple hepatitis B surface antigen epitopes during hepatitis B virus infection. J. Immunol.150:4659-4671.
9 Hwang, Y. K., N. K. Kim. J. M. Park. K. Lee. W. K. Han, H. I. Kim. and H. S.Cheong. 2002. HLA-A21 restricted peptides from the HBx antigen induce specific CTL responses in vitro and in vivo. Vaccine.20:3770-3777.
10 rtoletti a. Gehring AJ,2006. The immune response during hepatitis B virus infection.J Gen Virol.87:1439-1449.
11 Rehermann B, Fowler P'Sidney J. et al.1995. The cytotoxic T lymphocyte response to multiple hepatitis B virus polymerase epitopes during and after acute viral hepatitis. J Exp Med,181:1047-1058.
12 Kao, J.H., Chen, P.J., Lai, M.Y., Chen, D.S.2000.Hepatitis B genotypes correlate with clinical outcomes in patients with chronic hepatitis B. Gastroenterology.118:554-559.
13 C.H., Huang, C.C., Chen, W.J.2003. Prevalence of clinical implications of hepatitis B virus genotypes in southern Taiwan. Scand. J. Gastroenterol.1:95-101.
14 Lin, C.L., Kao, J.H.2011. The clinical implications of hepatitis B virus genotype:recent advances. J. Gastroenterol. Hepatol.26,123-130.
15 Hung CH, Chen CH, Lu SN, Wang JH, Hu TH, Huang CM, Tsai MC, Lee CM.2012. Precore/core promoter mutations and hepatitis B virus genotype in hepatitis B and C dually infected patients treated with interferon-based therapy. Antiviral Res.93 (1):55-63
16 Brusic V, Bajic VB, Petrovsky N.2004. Computational methods for prediction of T-cell epitopes-a framework for modelling, testing, and applications. Methods.34:436-443
17 Hironichi Tsurui, Takuya Takahashi. Prediction of T-cell epitope.2007. J Pharmacol Sci. 105:299-316.
18 Tan AT, Koh S, Goh W, Zhe HY, Gehring AJ, Lim SG, Bertoletti A.2010. A longitudinal analysis of innate and adaptive immune profile during hepatic flares in chronic hepatitis B. J Hepatol.52:330-339.
19 Vitiello A, Marchesini D, Furze J, Sherman LA, Chesnut RW.1991. Analysis of the HLA-restricted influenza-specific cytotoxic T lymphocyte response in transgenic mice carrying a chimeric human-mouse class Ⅰ major histocompatibility complex. J Exp Med. 173:1007-1015.
20 Chen A, Wang L, Zhang J, Zou L, Jia Z, Zhou W, et al.2005. H-2 Kd-restricted hepatitis B virus-derived epitope whose specific CD8+T lymphocytes can produce gamma interferon without cytotoxicity. J Virol.79(9): 5568-76.
21 吴玉章等.2002.针对病毒感染的治疗性疫苗研究评述.第三军医大学学报.Oct. Vol. 24, No.10.
22 Vitiello A, Ishioka G, Grey HM, Rose R, Farness P, LaFond R, Yuan L, Chisari FV, Furze J, Bartholomeuz R, et al.1995. Development of a lipopeptide-based therapeutic vaccine to treat chronic HBV infection. I. Induction of a primary cytotoxic T lymphocyte response in humans. J clin invest.95(1):341-349
23 Webster G, Bertoletti A.2001. Quantity and quality of virus-specific CD8 cell response: relevance to the design of a therapeutic vaccine for chronic HBV infection. Mol Immunol. 38(6)-467-473
24 Liu J, Zhang SH, Tan SG, Zheng BW, Gao GF.2011. Revival of the identification of cytotoxic T-lymphocyte epitopes for immunological diagnosis, therapy and vaccine development. Exp Biol Med.236:253-267.
25 Desmond CP, Bartholomeusz A, Gaudieri S, Re., Lewin SR.2008. A systematic review of T-cell epitopes in hepatitis B virus:identification, genotypic variation and relevance to antiviral therapeutics. Antivir Ther.13(2):161-175
1 Dunn C, Brunetto M, Reynolds G, Christophides T, Kennedy PT, Lampertico P, Das A, Lopes AR, Borrow P, Williams K, Humphreys E, Afford S, Adams DH, Bertoletti A, Maini MK.2007. Cytokines induced during chronic hepatitis B virus infection promote a pathway for NK cell-mediated liver damage. J Exp Med 204:667-680.
2 Zhang Z, Chen D, Yao J, Zhang H, Jin L, Shi M, Zhang H, Wang FS.2007. Increased infiltration of intrahepatic DC subsets closely correlate with viral control and liver injury in immune active pediatric patients with chronic hepatitis B. Clin Immunol 122:173-180.
3 Kakimi K, Lane TE, Wieland S, Asensio VC, Campbell IL, Chisari FV, Guidotti LG. 2001. Blocking chemokine responsive to gamma-2/interferon (IFN)-gamma inducible protein and monokine induced by IFN-gamma activity in vivo reduces the pathogenetic but not the antiviral potential of hepatitis B virus-specific cytotoxic T lymphocytes. J Exp Med 194:1755-1766.
4 Sitia G, Isogawa M, Kakimi K, Wieland SF, Chisari FV, Guidotti LG.2002. Depletion of neutrophils blocks the recruitment of antigen-nonspecific cells into the liver without affecting the antiviral activity of hepatitis B virus-specific cytotoxic T lymphocytes. Proc Natl Acad Sci U S A 99:13717-13722.
5 Stross L, Gunther J, Gasteiger G, Asen T, Graf S, Aichler M, Esposito I, Busch DH, Knolle P, Sparwasser T, Protzer U.2012. Foxp3+ regulatory T cells protect the liver from immune damage and compromise virus control during acute experimental hepatitis B virus infection in mice. Hepatology 56:873-883.
6 Zhang JY, Zhang Z. Lin F. Zou ZS. Xu RN. Jin L, Fu JL, Shi F. Shi M, Wang HF, Wang FS.2010. Interleukin-17-producing CD4(+) T cells increase with severity of liver damage in patients with chronic hepatitis B. Hepatology 51:81-91.
7 Leifeld L, Cheng S. Ramakers J, Dumoulin FL, Trautwein C, Sauerbruch T, Spengler U. 2002. Imbalanced intrahepatic expression of interleukin 12, interferon gamma, and interleukin 10 in fulminant hepatitis B. Hepatology 36:1001-1008.
8 Sitia G, Isogawa M, Iannacone M. Campbell IL. Chisari FV. Guidotti LG.2004. MMPs are required for recruitment of antigen-nonspecific mononuclcar cells into the liver by CTLs. J Clin Invest 113:1158-1167.
9 Tan AT. Koh S, Goh W, Zhe HY. Gehring AJ, Lim SG. Bertoletti A.2010. A longitudinal analysis of innate and adaptive immune profile during hepatic Hares in chronic hepatitis B. J Hcpatol 52:330-339.
10 Iannacone M, Sitia G, Ruggeri ZM, Guidotti LG.2007. HBV pathogenesis in animal models:recent advances on the role of platelets. J Hepatol 46:719-726.
11 Thimme R, Wieland S, Steiger C, Ghrayeb J, Reimann KA, Purcell RH, Chisari FV.2003. CD8(+) T cells mediate viral clearance and disease pathogenesis during acute hepatitis B virus infection.J Virol 77:68-76.
12 Maini MK, Boni C, Lee CK, Larrubia JR, Reignat S, Ogg GS, King AS, Herberg J, Gilson R, Alisa A, Williams R, Vergani D, Naoumov NV, Ferrari C, Bertoletti A.2000. 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.
13 Billioud G, Pichoud C, Parent R, Zoulim F.2012. Decreased infectivity of nucleoside analogs-resistant hepatitis B virus mutants. J Hepatol 56:1269-1275.
14 Amini-Bavil-Olyaee S, Herbers U, Sheldon J, Luedde T, Trautwein C, Tacke F.2009. The HA194T polymerase mutation impacts viral replication and susceptibility to tenofovir in hepatitis B e antigen-positive and hepatitis B e antigen-negative hepatitis B virus strains. Hepatology 49:1158-1165.
15 Sonneveld MJ, Rijckborst V, Zeuzem S, Heathcote EJ, Simon K, Senturk H, Pas SD, Hansen BE, Janssen HL.2012. Presence of precore and core promoter mutants limits the probability of respon.se to peginterferon in hepatitis B e antigen-positive chronic hepatitis B. Hepatology 56:67-75.
16 Yuen MF, Tanaka Y, Fong DY, Fung J, Wong DK, Yuen JC, But DY, Chan AO, Wong BC, Mizokami M, Lai CL.2009. Independent risk factors and predictive score for the development of hepatocellular carcinoma in chronic hepatitis B. J Hepatol 50:80-88.
17 Guo X, Jin Y, Qian G, Tu H.2008. Sequential accumulation of the mutations in core promoter of hepatitis B virus is associated with the development of hepatocellular carcinoma in Qidong, China. J Hepatol 49:718-725.
18 Xu Z, Ren X, Liu Y, Li X, Bai S, Zhong Y, Wang L, Mao P, Wang H, Xin S, Wong VW, Chan HL, Zoulim F, Xu D.2011. Association of hepatitis B virus mutations in basal core promoter and precore regions with severity of liver disease:an investigation of 793 Chinese patients with mild and severe chronic hepatitis B and acute-on-chronic liver failure..1 Gastroenterol 46:391-400.
19 Carey I, D'Antiga L, Bansal S, Longhi MS, Ma Y, Mesa IR. Mieli-Vergani G, Vergani D. 2011. Immune and viral profile from tolerance to hepatitis B surface antigen clearance:a longitudinal study of vertically hepatitis B virus-infected children on combined therapy. J Virol 85:2416-2428.
20 Abbott WGH, Tsai P, Leung E, Trevarton A, Ofanoa M, Homell J, Gane EJ, Munn SR, Rodrigo AG.2010. Associations between HLA Class I Alleles and Escape Mutations in the Hepatitis B Virus Core Gene in New Zealand-Resident Tongans. J Virol 84:621-629.
21 Maman Y, Blancher A, Benichou J, Yablonka A, Efroni S, Louzoun Y.2011. Immune-induced evolutionary selection focused on a single reading frame in overlapping hepatitis B virus proteins. J Virol 85:4558-4566.
22 Kim D, Lyoo KS, Smith D, Hur W, Hong SW, Sung PS, Yoon SK, Mehta S.2011. Number of Mutations Within CTL-Defined Epitopes of the Hepatitis B Virus (HBV) Core Region is Associated With HBV Disease Progression. J Med Virol 83:2082-2087.
23 Zhang XQ, Jiang L, You JP, Liu YY, Peng J, Zhang HY, et al.2011; Efficacy of short-term dexamethasone therapy in acute-on-chronic pre-liver failure. Hepatol Res. 41(1):46-53.
24 Vitiello A, Marchesini D, Furze J, Sherman LA, Chesnut RW.1991. Analysis of the HLA-restricted influenza-specific cytotoxic T lymphocyte response in transgenic mice carrying a chimeric human-mouse class I major histocompatibility complex. J Exp Med 173:1007-1015.
25 Chen A, Wang L, Zhang J, Zou L, Jia Z, Zhou W, et al. H-2 Kd-restricted hepatitis B virus-derived epitope whose specific CD8+T lymphocytes can produce gamma interferon without cytotoxicity. J Virol.2005; 79(9):5568-76.
26 Zlotnick A, Lee A, Bourne CR, Johnson JM, Domanico PL, Stray SJ.2007. In vitro screening for molecules that affect virus capsid assembly (and other protein association reactions). Nat Protoc 2:490-498.
27 Zlotnick A, Cheng N, Conway JF, Booy FP, Steven AC, Stahl SJ, Wingfield PT.1996. Dimorphism of hepatitis B virus capsids is strongly influenced by the C-terminus of the capsid protein. Biochemistry.35 (23):7412-21
28 Yang PL, Althage A, Chung J, Chisari FV.2002. Hydrodynamic injection of viral DNA: A mouse model of acute hepatitis B virus infection. Proc Natl Acad Sci U S A 99:13825-13830.
29 Desmond CP, Bartholomeusz A, Gaudieri S, Revill PA, Lewin SR.2008. A systematic review of T-cell epitopes in hepatitis B virus:identification. genotypic variation and relevance to antiviral therapeutics. AntivirTher 13:161-175.
30 Seeger. C. Mason.W.S..2000. Hepatitis B virus biology. Microbiol.Mol. Biol. Rev.64. 51-68.
31 Kim.W.H.. Honu. F.. Jaruga. B.. Hu.Z.. Fan. S.. Liang. T.J.. Gao. B..2001. Additive activation of hepatic NF-kappaB by ethanol and hepatitis B protein X (HBX) or HCV core protein:involvement of TNF-alpha receptor 1-independent and -dependent mechanisms. FASEB J.15,2551-2553.
32 Biermer, M., Puro, R., Schneider, R.J.,2003. Tumor necrosis factor alpha inhibition of hepatitis B virus replication involves disruption of capsid Integrity through activation i NF-kappaB. J. Virol.77,4033-4042.
33 Malik A, Singhal DK, Albanyan A, Husain SA, Kar P.2012. Hepatitis B virus gene mutations in liver diseases:a report from New Delhi. PloS one7:e39028.
34 Liu J, Zhang SH, Tan SG, Zheng BW, Gao GF.2011. Revival of the identification of cytotoxic T-lymphocyte epitopes for immunological diagnosis, therapy and vaccine development. Exp Biol Med 236:253-267.
35 Westover KM, Hughes AL.2007. Evolution of cytotoxic T-lymphocyte epitopes in hepatitis B virus. Infect Genet Evol 7:254-262.
36 Lai MW, Lin TY, Tsao KC, Huang CG, Hsiao MJ, Liang KH, Yeh CT.2012. Increased Seroprevalence of HBV DNA With Mutations in the S Gene Among Individuals Greater Than 18 Years Old After Complete Vaccination. Gastroenterology 143:400-407.
37 Frelin L, Wahlstrom T, Tucker AE, Jones J, Hughes J, Lee BO, Billaud JN, Peters C, Whitacre D, Peterson D, Milich DR.2009. A mechanism to explain the selection of the hepatitis e antigen-negative mutant during chronic hepatitis B virus infection. J Virol 83:1379-1392.
38 Wynne SA, Crowther RA, Leslie AG.1999. The crystal structure of the human hepatitis B virus capsid. Mol Cell 3:771-780.
39 Hosono S, Tai PC, Wang W, Ambrose M, Hwang DG, Yuan TT, Peng BH, Yang CS, Lee CS, Shih C.1995. Core antigen mutations of human hepatitis B virus in hepatomas accumulate in MHC class Ⅱ-restricted T cell epitopes. Virology.212:151-162.
40 Le Pogam S, Yuan TT, Sahu GK, Chatterjee S, Shih C.2000. Low-level secretion of human hepatitis B virus virions caused by two independent, naturally occurring mutations (P5T and L60V) in the capsid protein. J Virol 74:9099-9105
