Establishment of adeno-associated virus-hepatitis B virus mouse model

Abstract

Objective To establish an effective hepatitis B virus (HBV)-infected mouse model. Methods HEK293 cells were co-transfected with the three plasmids to produce adeno-associated virus (AAV)-HBV virus. After repeated freeze-thaw cycles, the virus was purified by density gradient centrifugation and identified. Twenty healthy adult C57BL/B6 mice were randomly divided into the study group (n=10) and the control group (n=10). The study group was injected with AAV-HBV virus through caudal vein, and the control group was injected with the same dose of AAV-GFP virus. The general condition in the two groups was determinted. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined at different time points. The quantitative detection of serum HBV DNA and HBsAg were examined by fluorescence quantitative PCR and chemiluminescence, respectively. After the mice were sacrificed, the livers were obtained for pathological examination. The HBsAg and HBcAg expression in the liver tissues were examined by immunohistochemistry. Results The purified AAV-HBV virus was successfully obtained with a purity of more than 97%. The general condition of mice infected with AAV - HBV virus was good with no abnormal behavior and normal liver function. No obvious pathological damage was observed in the liver tissues under the light microscope. The immunohistochemical staining showed positive HBsAg expression in the liver cytoplasm and positive HBcAg expression in the cell nucleus. The HBV DNA and HBsAg expression could be detected stably in the blood of mice infected with AAV-HBV virus. Conclusion The mouse model of HBV infection that is close to human beings has been established successfully, which can effectively and continuously produce virus. Key words: Adeno-associated virus; Hepatitis B virus; Disease model, animal; Mice