Construction and characterization of an infectious cDNA clone of rat hepatitis E virus

Rat hepatitis E virus (HEV) is related to human HEV and has been detected in wild rats worldwide. Here, the complete genome of rat HEV strain R63/DEU/2009 was cloned downstream of the T7 RNA polymerase promoter and capped genomic RNA generated by in vitro transcription was injected into nude rats. R...

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Published inJournal of general virology Vol. 96; no. Pt 6; pp. 1320 - 1327
Main Authors Li, Tian-Cheng, Yang, Tingting, Yoshizaki, Sayaka, Ami, Yasushi, Suzaki, Yuriko, Ishii, Koji, Haga, Kei, Nakamura, Tomofumi, Ochiai, Susumu, Takaji, Wakita, Johne, Reimar
Format Journal Article
LanguageEnglish
Published England 01.06.2015
Subjects
Animals
Cloning, Molecular - methods
DNA, Complementary - genetics
DNA, Viral - genetics
Feces - virology
Female
Hepatitis E virus - genetics
Hepatitis E virus - physiology
Injections, Intravenous
Male
Rats, Nude
Rats, Wistar
RNA, Viral - biosynthesis
RNA, Viral - genetics
Serum - virology
Transcription, Genetic
Virology - methods
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Summary:Rat hepatitis E virus (HEV) is related to human HEV and has been detected in wild rats worldwide. Here, the complete genome of rat HEV strain R63/DEU/2009 was cloned downstream of the T7 RNA polymerase promoter and capped genomic RNA generated by in vitro transcription was injected into nude rats. Rat HEV RNA could be detected in serum and faeces of rats injected intrahepatically, but not in those injected intravenously. Rat HEV RNA-positive faecal suspension was intravenously inoculated into nude rats and Wistar rats leading to rat HEV RNA detection in serum and faeces of nude rats, and to seroconversion in Wistar rats. In addition, rat HEV was isolated in PLC/PRF/5 cells from the rat HEV RNA-positive faecal suspension of nude rats and then passaged. The cell culture supernatant was infectious for nude rats. Genome analysis identified nine point mutations of the cell-culture-passaged virus in comparison with the originally cloned rat HEV genome. The results indicated that infectious rat HEV could be generated from the cDNA clone. As rats are widely used and well-characterized laboratory animals, studies on genetically engineered rat HEV may provide novel insights into organ tropism, replication and excretion kinetics as well as immunological changes induced by hepeviruses.
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ISSN:0022-1317
1465-2099
DOI:10.1099/vir.0.000072