Persistent growth of a human plasma-derived hepatitis C virus genotype 1b isolate in cell culture.
HCV (hepatitis C virus) research, including therapeutics and vaccine development, has been hampered by the lack of suitable tissue culture models. Development of cell culture systems for the growth of the most drug-resistant HCV genotype (1b) as well as natural isolates has remained a challenge. Transfection of cultured cells with adenovirus-associated RNA(I) (VA RNA(I)), a known interferon (IFN) antagonist and inhibitor of dsRNA-mediated antiviral pathways, enhanced the growth of plasma-derived HCV genotype 1b. Furthermore, persistent viral growth was achieved after passaging through IFN-alpha/beta-deficient VeroE6 cells for 2 years. Persistently infected cells were maintained in culture for an additional 4 years, and the virus rescued from these cells induced strong cytopathic effect (CPE). Using a CPE-based assay, we measured inhibition of viral production by anti-HCV specific inhibitors, including 2'-C-Methyl-D-Adenosine, demonstrating its utility for the evaluation of HCV antivirals. This virus constitutes a novel tool for the study of one of the most relevant strains of HCV, genotype 1b, which will now be available for HCV life cycle research and useful for the development of new therapeutics.
Silberstein
Erica
E
Division of Emerging and Transfusion-transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA.
Mihalik
Kathleen
K
Ulitzky
Laura
L
Plant
Ewan P
EP
Puig
Montserrat
M
Gagneten
Sara
S
Yu
Mei-ying W
MY
Kaushik-Basu
Neerja
N
Feinstone
Stephen M
SM
Taylor
Deborah R
DR
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D016923:Cell Death; D002522:Chlorocebus aethiops; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000254:growth & development* / Q000276:immunology; D006526:Hepatitis C / Q000097:blood / Q000188:drug therapy / Q000821:virology*; D018937:Hepatitis C Antibodies / Q000494:pharmacology; D018936:Hepatitis C Antigens / Q000235:genetics; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics; D016899:Interferon-beta / Q000235:genetics; D009500:Neutralization Tests; D020871:RNA Stability; D012367:RNA, Viral / Q000494:pharmacology; D014162:Transfection / Q000379:methods*; D014709:Vero Cells
D000998:Antiviral Agents; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D016898:Interferon-alpha; D012367:RNA, Viral; D016899:Interferon-beta