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Journal of Virology, May 2004, p. 4838-4846, Vol. 78, No. 9
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.9.4838-4846.2004

In Vitro Replication of Hepatitis E Virus (HEV) Genomes and of an HEV Replicon Expressing Green Fluorescent Protein

Suzanne U. Emerson,1* Hanh Nguyen,1 Judith Graff,1 David A. Stephany,2 Alicia Brockington,1 and Robert H. Purcell3

Molecular Hepatitis,1 Hepatitis Viruses Sections,3 Laboratory of Infectious Diseases and Flow Cytometry Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-80092

Received 19 August 2003/ Accepted 5 January 2004

Hepatitis E virus (HEV) RNA replication occurred in seven of nine primate cell cultures transfected with in vitro transcripts of an infectious cDNA clone. Cell-to-cell spread did not occur in cell cultures, but rhesus monkeys inoculated with lysates of HEV-transfected PLC/PRF/5 and Huh-7 cells became infected with HEV. A replicon with the ORF2 and ORF3 genes deleted and replaced with the green fluorescent protein gene also replicated in the same primate cells that supported the replication of the full-length genome. Fluorescence-activated cell sorter analysis confirmed that the 7mG cap structure was critical for efficient infectivity, although replication could be initiated at a very low level in its absence. HEV virions were also able to infect a limited number of cells of certain lines.

* Corresponding author. Mailing address: Bldg. 50, Room 6537, 50 South Dr., MSC-8009, Bethesda, MD 20892-8009. Phone: (301) 496-2787. Fax: (301) 402-0524. E-mail: semerson{at}niaid.nih.gov.

Journal of Virology, May 2004, p. 4838-4846, Vol. 78, No. 9
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.9.4838-4846.2004




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