| セッション情報 |
Enteric Viral Hepatitis i)Epidemiology of Hepatitis A and E
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| タイトル |
IF4a-3 Culture Systems for HEV
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| 演者 |
Hiroaki Okamoto（Division of VirologyDepartment of Infection and ImmunityJichi Medical University School of MedicineJapan） |
| 共同演者 |
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| 抄録 |
Early studies reported the propagation of hepatitis E virus （HEV） in either primary hepatocytes or severalestablishe・d cell linesbut replication was inefficient． Recentlyefficient culture systems have been estab－lished in PLC／PRF／5 （hepatocellular carcinoma） and A549 （lung cancer） cell lines for HEV strains of geno－types 3 and 4 in our laboratory． They originated in fecal extracts from patients containi／ng HEV RNA inextremely high－titers （107 copi’es／ml）and named the JEO3－1760F （genotype 3） and HE－JF5／15F （genotype4） strainsrespectively． HEV RNA in culture supernatants reached 108 copies／ml in titerand were trans－mitted successively through many passages． Various HEV strains with high roads of 1｝i 105 copies／ml incirculating blood from humanseven in fecesblood and liver homogenates from pigs and wlld boarswerealso propagated efficiently in PLC／PRF／5 and A549 cells． An infectious HEV cDNA clone （pJEO3－1760F／wt） was constructed that has replication activity comparable to that of the wild－type JEO3－1760F in feces．The ORF3 protein is indispensable for shedding HEV pa／rticlesfrom cells in the reverse genetics system．HEV recovered from culture mediaas well as circulating．肥Vpo．ssess ORF3 proteins o．血the surface andare covered with cellular membranesand thereforeORF2 epitopes are buried in these particles． In con－trastHEV excreted into feces are naked nucleocapsids without a lipid layer or surface expression of theORF3 protein． High－efficiency cell culture systems fOr infectious virusesthus developedare expected toopen up a new era and resolve many mysteries in the epidemiologymolecular biologyand treatment ofHEV． |
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