Development of plaque assays for hepatitis C virus-JFH1 strain and isolation of mutants with enhanced cytopathogenicity and replication capacity.

Abstract:

: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, GRP78 and phosphorylated eIF2-alpha, were overexpressed. Cells in the plaques were strongly positive for an apoptosis 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.

journal_name

Virology

journal_title

Virology

authors

Sekine-Osajima Y,Sakamoto N,Mishima K,Nakagawa M,Itsui Y,Tasaka M,Nishimura-Sakurai Y,Chen CH,Kanai T,Tsuchiya K,Wakita T,Enomoto N,Watanabe M

doi

10.1016/j.virol.2007.09.019

subject

Has Abstract

pub_date

2008-02-05 00:00:00

pages

71-85

issue

1

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(07)00597-1

journal_volume

371

pub_type

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