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The degree of susceptibility of human hepatoma (HepG2) cells to direct hepatitis B virus (HBV) infection remains unknown. We previously observed a low level of Dane particle production and viral DNA replication after in vitro infection of HepG2 cells with serum-derived HBV. However, this culture system appeared to be affected by variations as human hepatocyte cultures. In the present study, HBV infection of HepG2 cells led to a significant increase in the secretion of three envelope antigens (HBsAg, preS2Ag and preS1Ag) at 4 days post-infection, and Northern blot analysis revealed the presence of both preS1 (2.6 kb) and preS2/S (2.2 kb) transcripts. Expression of preS1Ag and the corresponding viral RNA became undetectable on 21 days post-infection whereas the 2.2 kb RNA species persisted and was associated with secretion of subviral HBs particles expressing preS2-epitopes and banding between 30 and 35% sucrose. At 35 days post-infection (fifth passage), a sudden high level production of HBsAg and preS1Ag was observed, followed by a massive cell death (90%). A stable HBsAg-producing HepG2 cell line, designated HepG2-BV3, grew out of the surviving cells. HepG2-BV3 cells could integrate HBV DNA sequences and produce the three HBV surface antigens. Treatment with dexamethasone increased the HBsAg and preS1Ag secretion. Such a HBsAg-producing HepG2 cell line obtained by in vitro HBV infection seems to mimick events that occur in the naturally occurring persistent chronic infection, and therefore may be an efficient in vitro model for studying the contribution of viral integration in the dysregulation of HBV and liver-specific genes expression.

journal of general virology

In vitro infection of human hepatoma cells (HepG2) with hepatitis B virus (HBV): spontaneous selection of a stable HBV surface antigen-producing HepG2 cell line containing integrated HBV DNA sequences