title: Response of hepatitis C virus to long-term passage in the presence of alpha interferon: multiple mutations and a common phenotype.
abstract: Cell culture-produced hepatitis C virus (HCV) has been subjected to up to 100 serial passages in human hepatoma cells in the absence or presence of different doses of alpha interferon (IFN-α). Virus survival, genetic changes, fitness levels, and phenotypic traits have been examined. While high initial IFN-α doses (increasing from 1 to 4 IU/ml) did not allow HCV survival beyond passage 40, a gradual exposure (from 0.25 to 10 IU/ml) allowed the virus to survive for at least 100 passages. The virus passaged in the presence of IFN-α acquired IFN-α resistance as evidenced by enhanced progeny production and viral protein expression in an IFN-α environment. A partial IFN-α resistance was also noted in populations passaged in the absence of IFN-α. All lineages acquired adaptative mutations, and multiple, nonsynonymous mutations scattered throughout the genome were present in IFN-α-selected populations. Comparison of consensus sequences indicates a dominance of synonymous versus nonsynonymous substitutions. IFN-α-resistant populations displayed decreased sensitivity to a combination of IFN-α and ribavirin. A phenotypic trait common to all assayed viral populations is the ability to increase shutoff host cell protein synthesis, accentuated in infections with IFN-α-selected populations carried out in the presence of IFN-α. The trait was associated with enhanced phosphorylation of protein kinase R (PKR) and eIF2α, although other contributing factors are likely. The results suggest that multiple, independent mutational pathways can confer IFN-α resistance to HCV and might explain why no unified picture has been obtained regarding IFN-α resistance in vivo.
authors: Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Consejo Superior de Investigaciones Científicas (CSIC), Campus de Cantoblanco, Madrid, Spain.
authors: Celia
authors: C
authors: Perales
authors: Nathan M
authors: NM
authors: Beach
authors: Isabel
authors: I
authors: Gallego
authors: Maria Eugenia
authors: ME
authors: Soria
authors: Josep
authors: J
authors: Quer
authors: Juan Ignacio
authors: JI
authors: Esteban
authors: Charles
authors: C
authors: Rice
authors: Esteban
authors: E
authors: Domingo
authors: Julie
authors: J
authors: Sheldon
mesh_terms: D000220:Adaptation, Biological / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000235:genetics; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation / Q000187:drug effects / Q000235:genetics; D010641:Phenotype; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012692:Serial Passage / Q000379:methods
chemical_list: D017931:DNA Primers; D016898:Interferon-alpha
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