title: Extinction of hepatitis C virus by ribavirin in hepatoma cells involves lethal mutagenesis.
abstract: Lethal mutagenesis, or virus extinction produced by enhanced mutation rates, is under investigation as an antiviral strategy that aims at counteracting the adaptive capacity of viral quasispecies, and avoiding selection of antiviral-escape mutants. To explore lethal mutagenesis of hepatitis C virus (HCV), it is important to establish whether ribavirin, the purine nucleoside analogue used in anti-HCV therapy, acts as a mutagenic agent during virus replication in cell culture. Here we report the effect of ribavirin during serial passages of HCV in human hepatoma Huh-7.5 cells, regarding viral progeny production and complexity of mutant spectra. Ribavirin produced an increase of mutant spectrum complexity and of the transition types associated with ribavirin mutagenesis, resulting in HCV extinction. Ribavirin-mediated depletion of intracellular GTP was not the major contributory factor to mutagenesis since mycophenolic acid evoked a similar decrease in GTP without an increase in mutant spectrum complexity. The intracellular concentration of the other nucleoside-triphosphates was elevated as a result of ribavirin treatment. Mycophenolic acid extinguished HCV without an intervening mutagenic activity. Ribavirin-mediated, but not mycophenolic acid-mediated, extinction of HCV occurred via a decrease of specific infectivity, a feature typical of lethal mutagenesis. We discuss some possibilities to explain disparate results on ribavirin mutagenesis of HCV.
authors: Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Consejo Superior de Investigaciones Científicas (CSIC), Campus de Cantoblanco, Madrid, Spain.
authors: Ana M
authors: AM
authors: Ortega-Prieto
authors: Julie
authors: J
authors: Sheldon
authors: Ana
authors: A
authors: Grande-Pérez
authors: Héctor
authors: H
authors: Tejero
authors: Josep
authors: J
authors: Gregori
authors: Josep
authors: J
authors: Quer
authors: Juan I
authors: JI
authors: Esteban
authors: Esteban
authors: E
authors: Domingo
authors: Celia
authors: C
authors: Perales
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D006160:Guanosine Triphosphate / Q000378:metabolism; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology / Q000821:virology; D006801:Humans; D020128:Inhibitory Concentration 50; D016296:Mutagenesis; D009153:Mutagens / Q000494:pharmacology*; D059645:Mutation Rate; D009173:Mycophenolic Acid / Q000494:pharmacology; D012367:RNA, Viral / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D012254:Ribavirin / Q000494:pharmacology*; D012692:Serial Passage
chemical_list: D000998:Antiviral Agents; D009153:Mutagens; D012367:RNA, Viral; D012254:Ribavirin; D006160:Guanosine Triphosphate; D009173:Mycophenolic Acid
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