Mutations That Permit Efficient Replication of Hepatitis C Virus RNA in Huh-7 Cells Prevent Productive Replication in Chimpanzees

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 99; no. 22; pp. 14416 - 14421
• Main Authors: Bukh, Jens, Pietschmann, Thomas, Lohmann, Volker, Krieger, Nicole, Faulk, Kristina, Engle, Ronald E., Govindarajan, Sugantha, Shapiro, Max, St. Claire, Marisa, Bartenschlager, Ralf
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 29.10.2002; National Acad Sciences
• Subjects: Adaptation, Physiological - genetics; Adaptation, Physiological - physiology; Amino acids; Animals; Biological Sciences; Cell culture techniques; Chimpanzees; Genetic mutation; Genome, Viral; Genomes; Hepacivirus - genetics; Hepacivirus - physiology; Hepatitis; Hepatitis C; Humans; Infections; Medical research; Mutagenesis; Mutation; Pan troglodytes; Replicon; Ribonucleic acid; RNA; RNA, Viral - biosynthesis; Transfection; Tumor Cells, Cultured; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - physiology; Virus Replication - genetics; Virus Replication - physiology; Viruses
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.212532699

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.