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

Research output: Contribution to journalArticle

55 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Hepatitis E virus
Macaca
Hepatocytes
Infection
Reverse Transcription
Hepatovirus
RNA
Myanmar
Serial Passage
Polymerase Chain Reaction
Antibodies
Liver
Serum-Free Culture Media
Viral RNA
Capsid Proteins
Collagenases
Virus Diseases
Virus Replication
Virion
Culture Media

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Tam, A. W., White, R., Yarbough, P. O., Murphy, B. J., McAtee, C. P., Lanford, R. E., & Fuerst, T. R. (1997). In vitro infection and replication of hepatitis E virus in primary cynomolgus macaque hepatocytes. Virology, 238(1), 94-102. https://doi.org/10.1006/viro.1997.8817

In vitro infection and replication of hepatitis E virus in primary cynomolgus macaque hepatocytes. / Tam, Albert W.; White, Robert; Yarbough, Patrice O.; Murphy, Brendan J.; McAtee, C. Patrick; Lanford, Robert E.; Fuerst, Thomas R.

In: Virology, Vol. 238, No. 1, 10.11.1997, p. 94-102.

Research output: Contribution to journalArticle

Tam, AW, White, R, Yarbough, PO, Murphy, BJ, McAtee, CP, Lanford, RE & Fuerst, TR 1997, 'In vitro infection and replication of hepatitis E virus in primary cynomolgus macaque hepatocytes', Virology, vol. 238, no. 1, pp. 94-102. https://doi.org/10.1006/viro.1997.8817
Tam AW, White R, Yarbough PO, Murphy BJ, McAtee CP, Lanford RE et al. In vitro infection and replication of hepatitis E virus in primary cynomolgus macaque hepatocytes. Virology. 1997 Nov 10;238(1):94-102. https://doi.org/10.1006/viro.1997.8817
Tam, Albert W. ; White, Robert ; Yarbough, Patrice O. ; Murphy, Brendan J. ; McAtee, C. Patrick ; Lanford, Robert E. ; Fuerst, Thomas R. / In vitro infection and replication of hepatitis E virus in primary cynomolgus macaque hepatocytes. In: Virology. 1997 ; Vol. 238, No. 1. pp. 94-102.
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