Production of infectious genotype 1a hepatitis C virus (Hutchinson strain) in cultured human hepatoma cells

Min Kyung Yi, Rodrigo A. Villanueva, David L. Thomas, Takaji Wakita, Stanley M. Lemon

Research output: Contribution to journalArticle

317 Scopus citations

Abstract

Infections with hepatitis C virus (HCV) are marked by frequent viral persistence, chronic liver disease, and extraordinary viral genetic diversity. Although much has been learned about HCV since its discovery, progress has been slowed by a lack of permissive cell culture systems supporting its replication. Productive infections have been achieved recently with genotype 2a virus, but cirrhosis and liver cancer are typically associated with genotype 1 HCV, which is more prevalent and relatively resistant to IFN therapy. We describe production of infectious genotype 1a HCV in cells transfected with synthetic RNA derived from a prototype virus (H77-S). Viral proteins accumulated more slowly in H77-S transfected cells than in cells transfected with genotype 2a (JFH-1) RNA, but substantially more H77-S RNA was secreted into supernatant fluids. Most secreted RNA was noninfectious, banding in isopycnic gradients at a density of 1.04-1.07 gm/cm3, but infectivity was associated with H77-S particles possessing a density of 1.13-1.14 gm/cm3. The specific infectivity of H77-S particles (5.4 × 104 RNA copies per focus-forming unit) was significantly lower than JFH-1 virus (1.4 × 102 RNA copies per focus-forming unit). Infection with either virus was blocked by CD81 antibody. Sera from genotype 1a-infected individuals neutralized H77-S virus, but had little activity against genotype 2a virus, suggesting that these genotypes represent different serotypes. The ability of this genotype 1a virus to infect cultured cells will substantially benefit antiviral and vaccine discovery programs.

Original languageEnglish (US)
Pages (from-to)2310-2315
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number7
DOIs
StatePublished - Feb 14 2006

Keywords

  • Bouyant density
  • CD81
  • Cell culture
  • Neutralizing antibody
  • Serotype

ASJC Scopus subject areas

  • General

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