Establishment of a mouse model stably replicating and expressing hepatitis B virus genotype C prevailed in China
10.11855/j.issn.0577-7402.2016.10.01
- Author:
Yue YANG
1
Author Information
1. Guilin Medical College
- Publication Type:Journal Article
- Keywords:
Animal;
Genotype;
Hepatitis B virus;
Models
- From:
Medical Journal of Chinese People's Liberation Army
2016;41(10):793-797
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a mouse model stably replicating and expressing hepatitis B virus genotype C (HBV-C) prevailed in China. Methods The recombinant adeno-associated virus rAAV8-1.3HBV-C (adr serotype) was transduced into the human hepatocellular carcinoma Huh7 cells in vitro, and the expression of HBsAg and HBeAg in the cell culture supernatant was determined by ELISA. High expression recombinant virus rAAV8-1.3HBV-C was screened and injected via the tail vein into eight C57BL/6 mice (aged 6-8 weeks) as the experimental group; meanwhile the previously reported rAAV8-1.3HBV-D (ayw serotype) was injected into seven C57BL/6 mice as the control group. HBV DNA load, HBsAg and HBeAg levels in sera were assayed at weeks 2, 3, 5, 7 and 9 post viral injections. Mice were sacrificed 9 weeks post injection and Hematoxylin-eosin (HE) staining and immunohistochemistry were performed to evaluate the pathological changes, and HBsAg and HBcAg expressions in the liver tissue. Results The supernatant HBsAg and HBeAg were detectable in the HuH7 cells 72h after transduction in vitro. The fluorescence quantitative PCR results of the HBV DNA load in serum at week 2, 3, 5, 7, and 9 post viral injection suggested stable in vivo replication of HBV DNA in mice. The serum expression of HBeAg was stable while the serum expression of HBsAg fluctuated. No obvious inflammatory cell infiltration or abnormal structure of liver tissue was observed, while HBsAg and HBcAg expression in the liver tissue were detected for both groups. Conclusion By in vivo transduction with the recombinant virus rAAV8-1.3HBV-C, a mouse model that stably expressed and replicated HBV-C has been successfully established.
