title: Stable Human Hepatoma Cell Lines for Efficient Regulated Expression of Nucleoside/Nucleotide Analog Resistant and Vaccine Escape Hepatitis B Virus Variants and Woolly Monkey Hepatitis B Virus.
abstract: Hepatitis B virus (HBV) causes acute and chronic hepatitis B (CHB). Due to its error-prone replication via reverse transcription, HBV can rapidly evolve variants that escape vaccination and/or become resistant to CHB treatment with nucleoside/nucleotide analogs (NAs). This is particularly problematic for the first generation NAs lamivudine and adefovir. Though now superseded by more potent NAs, both are still widely used. Furthermore, resistance against the older NAs can contribute to cross-resistance against more advanced NAs. For lack of feasible HBV infection systems, the biology of such variants is not well understood. From the recent discovery of Na+-taurocholate cotransporting polypeptide (NTCP) as an HBV receptor new in vitro infection systems are emerging, yet access to the required large amounts of virions, in particular variants, remains a limiting factor. Stably HBV producing cell lines address both issues by allowing to study intracellular viral replication and as a permanent source of defined virions. Accordingly, we generated a panel of new tetracycline regulated TetOFF HepG2 hepatoma cell lines which produce six lamivudine and adefovir resistance-associated and two vaccine escape variants of HBV as well as the model virus woolly monkey HBV (WMHBV). The cell line-borne viruses reproduced the expected NA resistance profiles and all were equally sensitive against a non-NA drug. The new cell lines should be valuable to investigate under standardized conditions HBV resistance and cross-resistance. With titers of secreted virions reaching >3 x 10(7) viral genome equivalents per ml they should also facilitate exploitation of the new in vitro infection systems.
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Cheng
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Guan
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.|Troop 66220 of PLA, Xingtai of Hebei Province, PR China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Sun
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Baosheng
authors.initials: B
authors.lastname: Li
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Haijun
authors.initials: H
authors.lastname: Li
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Fubiao
authors.initials: F
authors.lastname: Kang
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Jiwen
authors.initials: J
authors.lastname: Kang
authors.affiliation: Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China.
authors.forename: Dongliang
authors.initials: D
authors.lastname: Yang
authors.affiliation: Department of Internal Medicine II / Molecular Biology, University Hospital Freiburg, Freiburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Nassal
authors.affiliation: The Liver Disease Diagnosis and Treatment Center of PLA, Bethune International Peace Hospital, Shijiazhuang, PR China.
authors.forename: Dianxing
authors.initials: D
authors.lastname: Sun
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000821:virology*; D024882:Drug Resistance, Viral; D017325:Hepatitis B Vaccines / Q000008:administration & dosage; D006515:Hepatitis B virus / Q000187:drug effects* / Q000254:growth & development*; D019694:Hepatitis B, Chronic / Q000188:drug therapy / Q000821:virology*; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy / Q000821:virology; D014407:Tumor Cells, Cultured; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D017325:Hepatitis B Vaccines