title: Mutations that permit efficient replication of hepatitis C virus RNA in Huh-7 cells prevent productive replication in chimpanzees.
abstract: The development of a subgenomic replicon derived from the hepatitis C virus (HCV) strain Con1 enabled the study of viral RNA replication in Huh-7 cells. The level of replication of replicons, as well as full-length Con1 genomes, increased significantly by a combination of two adaptive mutations in NS3 (E1202G and T1280I) and a single mutation in NS5A (S2197P). However, these cell culture-adaptive mutations influenced in vivo infectivity. After intrahepatic transfection of chimpanzees, the wild-type Con1 genome was infectious and produced viral titers similar to those produced by other infectious HCV clones. Repeated independent transfections with RNA transcripts of a Con1 genome containing the three adaptive mutations failed to achieve active HCV infection. Furthermore, although a chimpanzee transfected with RNA transcripts of a Con1 genome with only the NS5A mutation became infected, this mutation was detected only in virus genomes recovered from serum at day 4; viruses recovered at day 7 had a reversion back to the original Con1 sequence. Our study demonstrates that mutations that are adaptive for replication of HCV in cell culture may be highly attenuating in vivo.
authors.affiliation: Hepatitis Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA. jbukh@niaid.nih.gov
authors.forename: Jens
authors.initials: J
authors.lastname: Bukh
authors.forename: Thomas
authors.initials: T
authors.lastname: Pietschmann
authors.forename: Volker
authors.initials: V
authors.lastname: Lohmann
authors.forename: Nicole
authors.initials: N
authors.lastname: Krieger
authors.forename: Kristina
authors.initials: K
authors.lastname: Faulk
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Engle
authors.forename: Sugantha
authors.initials: S
authors.lastname: Govindarajan
authors.forename: Max
authors.initials: M
authors.lastname: Shapiro
authors.forename: Marisa
authors.initials: M
authors.lastname: St  Claire
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics* / Q000502:physiology; D000818:Animals; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D016296:Mutagenesis; D002679:Pan troglodytes; D012367:RNA, Viral / Q000096:biosynthesis; D012093:Replicon; D014407:Tumor Cells, Cultured; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000502:physiology; D014779:Virus Replication / Q000235:genetics* / Q000502:physiology; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
chemical_list: D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase