The dose of HBV genome contained plasmid has a great impact on HBV persistence in hydrodynamic injection mouse model.

ABSTRACT: Hydrodynamic injection (HI) of hepatitis B virus (HBV) mouse model is an useful tool for HBV related research in vivo. However, only 40% of C57/BL6 mice injected with 10 ?g HBV genome contained plasmid (pAAV-HBV1.2), serum HBsAg more than 6 months and none of the BALB/c mice injected with 10 ?g pAAV-HBV1.2 plasmid DNA, serum HBsAg positive more than 4 weeks in the previous study.In this study, C57/BL6 and BALB/c mice were hydrodynamic injected with different doses of pAAV-HBV1.2 plasmid DNA. HBV related serum markers were detected by ELISA. ALT levels in the serum were measured using full automated biochemistry analyzer. HBcAg positive cells in the liver were detected by immunohistochemical staining. The mRNA levels of IRF3, ISGs including ISG15, OAS, PKR and immune factors including IFN?, TNF?, TGF?, IL-6, IL-10, PDL1 in liver of the mice were quantified by qRT-PCR.The results showed that the mice injected with 100 ?g high-concentration or 1 ?g low-concentration of pAAV-HBV1.2 plasmid DNA did not excert dominant influence on HBV persistence. In contrast, injection of 5 ?g intermediate-dose of pAAV-HBV1.2 plasmid DNA led to significant prolonged HBsAg expression and HBV persistence in both C57/BL6 (80% of the mice with HBsAg positive more than 6 months) and BALB/c (60% of the mice with HBsAg positive more than 3 months) mice. IFN? was significant up-regulated in liver of the mice injected with 1 ?g or 100 ?g pAAV-HBV1.2 plasmid DNA. TNF? was up-regulated significantly in liver of the mice injected with 100 ?g pAAV-HBV1.2 plasmid DNA. Moreover, PDL1 was significant up-regulated in liver of the mice injected with 5 ?g pAAV-HBV1.2 plasmid DNA.In this paper we demonstrated that, in the HBV HI mouse model, the concentration of injected pAAV-HBV1.2 plasmid DNA contributes to the diverse kinetics of HBsAg and HBeAg in the serum as well as HBcAg expression level in the liver, which then determined the HBV persisternce, while the antiviral factors IFN?, TNF? as well as immune negative regulatory factor PDL1 play important roles on HBV persistence.

SUBMITTER: Li L 

PROVIDER: S-EPMC5657044 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature