J Virol. 2018 Sep 19. pii: JVI.01255-18. doi: 10.1128/JVI.01255-18. [Epub ahead of print]
Robust human and murine hepatocyte culture models of hepatitis B virus infection and replication.
Qiao L1, Sui J2, Luo G3.
Author information
1
Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama, USA.
2
National Institute of Biological Sciences, Beijing, China.
3
Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama, USA [email protected].
Abstract
Hepatitis B virus (HBV) is a major cause of chronic liver diseases including hepatitis, cirrhosis, and hepatocellular carcinoma. HBV research has been hampered by the lack of robust cell culture and small animal models of HBV infection. The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as an HBV receptor has been a landmark advance in HBV research in recent years. Ectopic expression of NTCP in nonpermissive HepG2, Huh7, and AML12 cell lines confers HBV susceptibility. However, HBV replication in these human and murine hepatocytes cell lines appeared suboptimal. In present study, we have constructed stable NTCP-expressing HepG2 and AML12 cell lines and found that HBV permissiveness is correlated with NTCP expression. More significantly, we have developed robust HBV cell culture models by treating the HBV-infected cells with dimethyl sulfoxide (DMSO) and hydrocortisone, which significantly promoted HBV replication and production. Mechanistic studies suggested that hydrocortisone significantly enhanced the transcription and expression of PGC1α and HNF4α which are known to promote HBV transcription and replication. These new human and murine hepatocyte culture systems of HBV infection and replication will accelerate the determination of molecular aspects underlying HBV infection, replication, and morphogenesis in human and murine hepatocytes. We anticipate that our HBV cell culture models will also facilitate the discovery and development of antiviral drugs towards the ultimate eradication of chronic hepatitis B.IMPORTANCEHBV research has been greatly hampered by the lack of robust cell culture and small animal models of HBV infection and propagation. The discovery of NTCP as an HBV receptor has greatly impacted the field of HBV research. Although HBV infection of the NTCP-expressing human and murine hepatocyte cell lines has been demonstrated, its replication in cell culture appeared inefficient. To further improve cell culture systems of HBV infection and replication, we have constructed NTCP-expressing HepG2 and AML-12 cell lines that are highly permissive to HBV infection. More significantly, we have found that DMSO and hydrocortisone markedly enhanced HBV transcription and replication in human and murine hepatocytes when added to cell culture medium. These new cell culture models of HBV infection and replication will facilitate HBV research and antiviral drug discovery towards ultimate elimination of chronic hepatitis B.