In vitro infection and replication of hepatitis E virus in primary cynomolgus macaque hepatocytes

Albert W. Tam, Robert White, Patrice O. Yarbough, Brendan J. Murphy, C. Patrick McAtee, Robert E. Lanford, Thomas R. Fuerst

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    59 Scopus citations

    Abstract

    An in vitro model was developed to replicate hepatitis E virus (HEV) in normal primary cynomolgus macaque hepatocytes using a hormonally defined, serum-free medium formulation. Primary hepatocytes were infected in tissue culture following isolation by collagenase treatment of liver wedge biopsy material. Viral replication was monitored by a highly strand-specific reverse transcription-polymerase chain reaction (RT-PCR) assay, which could detect the positive- and negative-strands of HEV RNA independently in a sensitive and specific manner. Several infectious HEV (Burma strain) inocula were titered by this RT-PCR assay, and a minimum effective infectious dose was determined. Appearance of newly replicated virus was demonstrated by detection of both strands of HEV RNA in experimentally infected hepatocytes as well as the genomic positive-strand viral RNA in the culture medium. Infectivity of the virus particles present in the media was confirmed by serial passage and replication of the virus in culture. Using this in vitro infection system, a neutralization assay was developed to assess the ability of anti-HEV antibodies to block virus infection of liver cells. Results presented in this report represent the first in vitro demonstration of a neutralizing anti-HEV antibody directed against the ORF2-encoded putative capsid protein.

    Original languageEnglish (US)
    Pages (from-to)94-102
    Number of pages9
    JournalVirology
    Volume238
    Issue number1
    DOIs
    StatePublished - Nov 10 1997

    ASJC Scopus subject areas

    • Virology

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