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Regulation by dimethylsulfoxide, insulin, and corticosteroids of hepatitis b virus replication in a transfected human hepatoma cell line
Author(s) -
Gripon Philippe,
Diot Christian,
Corlu Anne,
GuguenGuillouzo Christiane
Publication year - 1989
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890280316
Subject(s) - transfection , viral replication , biology , cell culture , virology , clone (java method) , hepatitis b virus , intracellular , microbiology and biotechnology , virus , extracellular , hepatocyte , dna , plasmid , hepatitis b virus pre beta , in vitro , hepatitis b virus dna polymerase , biochemistry , genetics
A human hepatoblastoma cell clone E4 was obtained by transfection of HepG 2 cells with a plasmid DNA containing four tandem copies of hepatitis B virus (HBV) genome. Analysis of both intracellular and extracellular viral DNA revealed that this clone exhibited the main steps of the replication process previously found in normal hepatocyte primary cultures experimentally infected in vitro. Indeed, relaxed‐circular, covalently closed circular, and single‐stranded forms of viral DNA were identified in the cells together with complete virions and immature cores in the medium. Furthermore, the ability of these secreted particles to infect normal human hepatocyte cultures was established. These E4 cells were used to evaluate the effect of various soluble factors on HBV replication. Corticosteroids and, to a greater extent, dimethylsulfoxide (DMSO) increased intracellular viral DNA, whereas insulin reduced it dramatically. Parallel changes in the amounts of viral DNA secreted in the medium were observed. Measurement of the albumin secretion rate indicated that cellular and viral activities could be regulated, at least in part, in a coordinated manner.