Demonstration of in Vitro Infection of Chimpanzee Hepatocytes with Hepatitis C Virus Using Strand-Specific RT/PCR

Robert E. Lanford, Camille Sureau, James R. Jacob, Robert White, Thomas R. Fuerst

Research output: Contribution to journalArticlepeer-review

278 Scopus citations

Abstract

Analysis of hepatitis C virus (HCV) replication has been hampered due to the difficulty encountered in in vitro cultivation of the virus in conventional tissue culture systems. In this study, primary chimpanzee hepatocyte cultures maintained in a serum-free medium formulation were susceptible to in vitro infection with HCV. In order to document infection, two new methods of reverse transcription/polymerase chain reaction were developed that permit accurate distinction between positive and negative strand HCV RNA. One method relied upon the use of a tagged cDNA primer, while the second method employed a thermostable reverse transcriptase. Following inoculation of chimpanzee hepatocytes with HCV, intracellular positive and negative strand HCV RNA were detectable 4 days postinfection and throughout the remainder of the experimental period, 25 days. Analysis of HCV-inoculated baboon hepatocytes revealed a total absence of negative strand HCV RNA, while residual positive strand RNA from the inoculum could be detected for up to 11 days. The in vitro replication of HCV RNA in chimpanzee hepatocytes could be suppressed by α-interferon. This system should be amenable to the study of HCV replication, antiviral compounds, and the development of neutralization assays.

Original languageEnglish (US)
Article number71381
Pages (from-to)606-614
Number of pages9
JournalVirology
Volume202
Issue number2
DOIs
StatePublished - Aug 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Virology

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