Viral evolution and interferon resistance of hepatitis C virus RNA replication in a cell culture model

Rhea Sumpter, Chunfu Wang, Eileen Foy, Yueh Ming Loo, Michael Gale

Research output: Contribution to journalArticle

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Abstract

Hepatitis C virus (HCV) replicates through an error-prone process that may support the evolution of genetic variants resistant to the host cell antiviral response and interferon (IFN)-based therapy. We evaluated HCV-IFN interactions within a long-term culture system of Huh7 cell lines harboring different variants of an HCV type 1b subgenomic RNA replicon that differed at only two sites within the NS5A-encoding region. A replicon with a K insertion at HCV codon 2040 replicated efficiently and exhibited sequence stability in the absence of host antiviral pressure. In contrast, a replicon with an L2198S point mutation replicated poorly and triggered a cellular response characterized by EFN-β production and low-level IFN-stimulated gene (ISG) expression. When maintained in long term-culture, the L2198S RNA evolved into a stable high-passage (HP) variant with six additional point mutations throughout the HCV protein-encoding region that enhanced viral replication. The HP RNA transduced Huh7 cells with more than 1,000-fold greater efficiency than its L2198S progenitor or the K2040 sequence. Replication of the HP RNA resisted suppression by IFN-α treatment and was associated with virus-directed reduction in host cell expression of ISG56, an antagonist of HCV RNA translation. Accordingly, the HP RNA was retained within polyribosome complexes in vivo that were refractory to IFN-induced disassembly. These results identify ISG56 as a translational control effector of the host response to HCV and provide direct evidence to link this response to viral sequence evolution, ISG regulation, and selection of the IFN-resistant viral phenotype.

Original languageEnglish (US)
Pages (from-to)11591-11604
Number of pages14
JournalJournal of Virology
Volume78
Issue number21
DOIs
StatePublished - Nov 2004

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Hepatitis C virus
interferons
Virus Replication
Hepacivirus
Interferons
cell culture
Cell Culture Techniques
RNA
replicon
Replicon
point mutation
Point Mutation
Antiviral Agents
Satellite Viruses
polyribosomes
Polyribosomes
Molecular Evolution
cells
virus replication
RNA replication

ASJC Scopus subject areas

  • Immunology

Cite this

Viral evolution and interferon resistance of hepatitis C virus RNA replication in a cell culture model. / Sumpter, Rhea; Wang, Chunfu; Foy, Eileen; Loo, Yueh Ming; Gale, Michael.

In: Journal of Virology, Vol. 78, No. 21, 11.2004, p. 11591-11604.

Research output: Contribution to journalArticle

Sumpter, Rhea ; Wang, Chunfu ; Foy, Eileen ; Loo, Yueh Ming ; Gale, Michael. / Viral evolution and interferon resistance of hepatitis C virus RNA replication in a cell culture model. In: Journal of Virology. 2004 ; Vol. 78, No. 21. pp. 11591-11604.
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