Journal
VIROLOGY
Volume 371, Issue 1, Pages 71-85Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2007.09.019
Keywords
HCV-JFH1; HCV cell culture; plaque assay; ER stress; unfolded protein responses; apoptosis; NS5B RNA-dependent RNA polymerase
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HCV culture in vitro results in massive cell death, which suggests the presence of HCV-induced cytopathic effects. Therefore, we investigated its mechanisms and viral nucleotide sequences involved in this effect using HCV-JFH1 cell culture and a newly developed HCV plaque assay technique. The plaque assay developed cytopathic plaques, depending on the titer of the inoculum. In the virus-infected cells, the ER stress markers, GR-P78 and phosphorylated eIF2-alpha, were overexpressed. Cells in the plaques were strongly positive for an apoplosis marker, annexin V Isolated virus subclones from individual plaque showed greater replication efficiency and cytopathogenicity than the parental virus. The plaque-purified virus had 9 amino acid substitutions, of which 5 were clustered in the C terminal of the NS5B region. Taken together, the cytopathic effect of HCV infection involves ER-stress-induced apoptotic cell death. Certain HCV genomic structures may determine the viral replication capacity and cytopathogenicity. (c) 2007 Elsevier Inc. All rights reserved.
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