Establishment and preliminary evaluation of HBsAg mutated hepatitis B transfected mice model via hydrodynamic injection

Abstract

Abstract: Objective To investigate the role of hepatitis B surface antigen (HBsAg) in chronicity of hepatitis B virus (HBV) and regulation of host immune responses in vivo.Methods Site-directed mutagenesis was used to establish HBsAg mutated HBV mice model, based on transfection of plasmid adeno-associated virus (pAAV)-HBV through hydrodynamic injection. Then the serum HBsAg, hepatitis B e antigen (HBeAg) and HBV DNA were measured to evaluate the infection in both mice models. In addition, hematoxylin-eosin (HE) staining was performed to analyze pathological changes and infiltration of immune cells in liver.Results HBsAg mutated HBV mice model was successfully established. HBeAg levels in HBV mice model and HBsAg mutated HBV mice model were both 4 COI/ml with no difference. HBsAg and HBV DNA were undetected in HBsAg mutated HBV mice model, while HBsAg was > 10² COI/ml and HBV DNA was 10³ ~ 10⁶ copies/ml in HBV mice model. Dynamic monitor showed that percentage of HBeAg positive rate gradually declined in both mice models, which was more obvious in HBsAg mutated HBV model. At week 8, HBeAg positive rate was 20% in HBsAg mutated HBV model compared to 60% in HBV model. Compared with HBV model, there was less infiltration of immune cells but more ground-glass hepatocytes were in the liver of HBsAg mutated HBV mice model with HBsAg deficiency. Conclusion HBsAg mutated HBV mice model was successfully established. HBsAg might have an important role in pathologic process and infiltration of immune cells in liver. It lays a foundation for further study on the role of HBsAg in HBV replication and immunologic escape.

Key words: Chronic hepatitis B, HBV, HBsAg, Hydrodynamic injection, Ground-glass appearance, Infiltration of immune cells

ZHANG Yi, CHEN Jie-liang, FANG Zhong, XU Guo-guang, LI Jun.. Establishment and preliminary evaluation of HBsAg mutated hepatitis B transfected mice model via hydrodynamic injection[J]. Chinese Hepatolgy, 2017, 22(10): 894-898.

References

[1] Ganem D, Prince AM. Hepatitis B virus infection--natural history and clinical consequences. N Engl J Med, 2004, 350: 1118-1129.
[2] Dienstag JL. Hepatitis B virus infection. N Engl J Med, 2008, 359: 1486-1500.
[3] Wang S, Chen Z, Hu C, et al. Hepatitis B virus surface antigen selectively inhibits TLR2 ligand-induced IL-12 production in monocytes/macrophages by interfering with JNK activation. J Immunol, 2013, 190: 5142-5151.
[4] Kosinska AD, Pishraft-Sabet L, Wu W, et al. Low hepatitis B virus-specific T-cell response in males correlates with high regulatory T-cell numbers in murine models. Hepatology, 2017, 66:69-83.
[5] Fang Z, Li J, Yu X, et al. Polarization of monocytic myeloid-derived suppressor cells by hepatitis B surface antigen is mediated via ERK/IL-6/STAT3 signaling feedback and restrains the activation of T cells in chronic hepatitis B virus infection. J Immunol, 2015, 195: 4873-4883.
[6] Loggi E, Bihl FK, Cursaro C, et al. Virus-specific immune response in HBeAg-negative chronic hepatitis B: relationship with clinical profile and HBsAg serum levels. PLoS One, 2013, 8: e65327.
[7] Skrastina D, Bulavaite A, Sominskaya I, et al. High immunogenicity of a hydrophilic component of the hepatitis B virus preS1 sequence exposed on the surface of three virus-like particle carriers. Vaccine, 2008, 26: 1972-1981.
[8] Hruska JF, Robinson WS. The proteins of hepatitis B Dane particle cores. J Med Virol, 1977, 1: 119-131.
[9] Yang PL, Althage A, Chung J, et al. Hydrodynamic injection of viral DNA: a mouse model of acute hepatitis B virus infection. Proc Natl Acad Sci U S A, 2002, 99: 13825-13830.
[10] Chisari FV. Hepatitis B virus transgenic mice: models of viral immunobiology and pathogenesis. Curr Top Microbiol Immunol, 1996, 206: 149-173.
[11] Guidotti LG, Chisari FV. Immunobiology and pathogenesis of viral hepatitis. Annu Rev Pathol, 2006, 1: 23-61.
[12] Chuai X, Wang W, Chen H, et al. Lentiviral backbone-based hepatitis B virus replicon-mediated transfer favours the establishment of persistent hepatitis B virus infection in mice after hydrodynamic injection. Antiviral Res, 2014, 101: 68-74.
[13] Liang TJ. Hepatitis B: the virus and disease. Hepatology, 2009, 49: S13-S21.
[14] Glebe D, Urban S. Viral and cellular determinants involved in hepadnaviral entry. World J Gastroenterol, 2007, 13: 22-38.
[15] Yan H, Zhong G, Xu G, et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife, 2012, 1: e49.
[16] Wu PC, Lam KC. Cytoplasmic hepatitis B surface antigen and the ground-glass appearance in hepatocellular carcinoma. Am J Clin Pathol, 1979, 71: 229-234.
[17] Reifenberg K, Hildt E, Lecher B, et al. IFNgamma expression inhibits LHBs storage disease and ground glass hepatocyte appearance, but exacerbates inflammation and apoptosis in HBV surface protein-accumulating transgenic livers. Liver Int, 2006, 26: 986-993.