Abstract
BackgroundHydrodynamic 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 μg HBV genome contained plasmid (pAAV-HBV1.2), serum HBsAg more than 6 months and none of the BALB/c mice injected with 10 μg pAAV-HBV1.2 plasmid DNA, serum HBsAg positive more than 4 weeks in the previous study.MethodsIn 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 IFNγ, TNFα, TGFβ, IL-6, IL-10, PDL1 in liver of the mice were quantified by qRT-PCR.ResultsThe results showed that the mice injected with 100 μg high-concentration or 1 μg low-concentration of pAAV-HBV1.2 plasmid DNA did not excert dominant influence on HBV persistence. In contrast, injection of 5 μg 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. IFNγ was significant up-regulated in liver of the mice injected with 1 μg or 100 μg pAAV-HBV1.2 plasmid DNA. TNFα was up-regulated significantly in liver of the mice injected with 100 μg pAAV-HBV1.2 plasmid DNA. Moreover, PDL1 was significant up-regulated in liver of the mice injected with 5 μg pAAV-HBV1.2 plasmid DNA.ConclusionIn 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 IFNγ, TNFα as well as immune negative regulatory factor PDL1 play important roles on HBV persistence.
Highlights
Hydrodynamic injection (HI) of hepatitis B virus (HBV) mouse model is an useful tool for HBV related research in vivo
We found that the antiviral factor IFNγ was significant up-regulated in liver of the mice injected with 1 μg or 100 μg pAAVHBV1.2 plasmid DNA
Kinetics of HBV surface antigenemia (HBsAg), HBeAg, HBcAb, HBsAb in sera of both C57/BL6 and BALB/c mice after HI To investigate whether the dose of injected HBV plasmids has a great impact on HBV persistence, 5 μg, 10 μg or 100 μg pAAV-HBV1.2 plasmid DNA were injected into the C57/BL6 mice, respectively by HI method
Summary
Hydrodynamic injection (HI) of hepatitis B virus (HBV) mouse model is an useful tool for HBV related research in vivo. An immunologically defined and reproducible small animal model for HBV infection remains unavailable due to the strict host specificity of HBV infection, which greatly hampers HBV related research. Several lines of transgenic mice with replication competent HBV genomes have been established and showed powerful application value for HBV related research [6]. Transgenic mice have integrated HBV genome and HBV replication existed in all the hepatocytes. The presence of HBV genomes in these mouse lines inevitably induces host immune tolerance against HBV antigens, which is different from that occurs during a natural infection [1, 6, 9, 15]. The transplant models are based on immunodeficient mouse strains and difficult to operate in majority of laboratory
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