In vitro replication of hepatitis E virus (HEV) genomes and of an HEV replicon expressing green fluorescent protein.
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.
Emerson
Suzanne U
SU
Laboratory of Infectious Diseases and Flow Cytometry Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-8009, USA. semerson@niaid.nih.gov
Nguyen
Hanh
H
Graff
Judith
J
Stephany
David A
DA
Brockington
Alicia
A
Purcell
Robert H
RH
D000818:Animals; D002460:Cell Line; D018076:DNA, Complementary; D016679:Genome, Viral; D049452:Green Fluorescent Proteins; D016751:Hepatitis E / Q000503:physiopathology / Q000821:virology; D016752:Hepatitis E virus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D008253:Macaca mulatta; D011323:Primates; D012093:Replicon; D014162:Transfection; D014771:Virion / Q000378:metabolism; D014779:Virus Replication
D018076:DNA, Complementary; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins