Control of Hepatitis B Virus Replication by Innate Response of HepaRG Cells

Hepatitis B virus (HBV) is currently viewed as a stealth virus that does not elicit innate immunity in vivo. This assumption has not yet been challenged in vitro because of the lack of a relevant cell culture system. The HepaRG cell line, which is physiologically closer to differentiated hepatocytes and permissive to HBV infection, has opened new perspectives in this respect. HBV baculoviruses were used to initiate an HBV replication in both HepG2 and HepaRG cells. To monitor HBV replication, the synthesis of encapsidated DNA, and secretion of hepatitis B surface antigen (HBsAg), was respectively analyzed by southern blot and enzyme-linked immunosorbent assay. The induction of a type I interferon (IFN) response was monitored by targeted quantitative reverse transcription polymerase chain reaction (qRT-PCR), low-density arrays, and functional assays. The invalidation of type I IFN response was obtained by either antibody neutralization or RNA interference. We demonstrate that HBV elicits a strong and specific innate antiviral response that results in a noncytopathic clearance of HBV DNA in HepaRG cells. Challenge experiment showed that transduction with Bac-HBV-WT, but not with control baculoviruses, leads to this antiviral response in HepaRG cells, whereas no antiviral response is observed in HepG2 cells. Cellular gene expression analyses showed that IFN-beta and other IFN-stimulated genes were up-regulated in HepG2 and HepaRG cells, but not in cells transduced by control baculoviruses. Interestingly, a rescue of viral replication was observed when IFN-beta action was neutralized by antibodies or RNA interference of type I IFN receptor. Conclusion: Our data suggest that a strong HBV replication is able to elicit a type I IFN response in HepaRG-transduced cells. (HEPATOLOGY 2010;51:63-72.)