In vitro propagation and production of hepatitis E virus from in vivo-infected primary macaque hepatocytes

Albert W. Tam, Robert White, Edith Reed, Mary Short, Yifan Zhang, Thomas R. Fuerst, Robert E. Lanford

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    57 Scopus citations

    Abstract

    Hepatitis E virus (HEV) is responsible for sporadic cases as well as large epidemics of acute viral hepatitis in many developing countries. The nucleotide sequence of HEV appears to be unique among known viruses and thus may represent a prototype human pathogen in a novel class of single-stranded, positive-sense RNA viruses. To facilitate further studies of the biology of HEV, a tissue culture system using a serum-free medium formulation has been developed to propagate the virus in vitro. Hepatocytes were isolated from livers of cynomolgus macaques experimentally infected with a HEV (Burma strain) inoculum and maintained in long-term cultures. Using a highly strand-specific RT-PCR assay, both the positive-sense and the negative replicative strands of HEV RNA were detected in these hepatocytes throughout the course of the experiments. Positive-strand genomic RNA was also detected in the culture medium, suggesting the production and secretion of HEV virus particles. The virus particles were successfully concentrated 200-fold from the medium using ultrafiltration, and they could be observed by immunoelectron microscopy using anti-HEV-positive immune serum. These results demonstrate the capacity of this hepatocyte culture system to replicate HEV in vitro, thus providing an experimental means to study the replicative process of the virus.

    Original languageEnglish (US)
    Pages (from-to)1-9
    Number of pages9
    JournalVirology
    Volume215
    Issue number1
    DOIs
    StatePublished - Jan 1 1996

    ASJC Scopus subject areas

    • Virology

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  • Cite this

    Tam, A. W., White, R., Reed, E., Short, M., Zhang, Y., Fuerst, T. R., & Lanford, R. E. (1996). In vitro propagation and production of hepatitis E virus from in vivo-infected primary macaque hepatocytes. Virology, 215(1), 1-9. https://doi.org/10.1006/viro.1996.0001