title: Rat hepatitis E virus derived from wild rats (Rattus rattus) propagates efficiently in human hepatoma cell lines.
abstract: Although rat hepatitis E virus (HEV) has been identified in wild rats, no cell culture systems for this virus have been established. A recent report suggesting the presence of antibodies against rat HEV in human sera encouraged us to cultivate rat HEV in human cells. When liver homogenates obtained from wild rats (Rattus rattus) in Indonesia were inoculated onto human hepatocarcinoma cells, the rat HEV replicated efficiently in PLC/PRF/5, HuH-7 and HepG2 cells, irrespective of its genetic group (G1-G3). The rat HEV particles released from cultured cells harbored lipid-associated membranes on their surface that were depleted by treatment with detergent and protease, with the buoyant density in sucrose shifting from 1.15-1.16 g/ml to 1.27-1.28 g/ml. A Northern blotting analysis revealed genomic RNA of 7.0 kb and subgenomic RNA of 2.0 kb in the infected cells. The subgenomic RNA of G1-G3 each possessed the extreme 5'-end sequence of GUAGC (nt 4933-4937), downstream of the highly conserved sequence of GAAUAACA (nt 4916-4923). The establishment of culture systems for rat HEV would allow for extended studies of the mechanisms of viral replication and functional roles of HEV proteins. Further investigation is required to clarify the zoonotic potential of rat HEV.
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan.
authors.forename: Suljid
authors.initials: S
authors.lastname: Jirintai
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan.
authors.lastname: Tanggis
authors.affiliation: West Nusa Tenggara Hepatitis Laboratory, Mataram, Indonesia; Immunobiology Laboratory, Faculty of Medicine, University of Mataram, Mataram, Indonesia.
authors.lastname: Mulyanto
authors.affiliation: Department of Clinical Pathology, Moewardi Hospital, University of Sebelas Maret, Solo, Indonesia.
authors.forename: Joseph Benedictus
authors.initials: JB
authors.lastname: Suparyatmo
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan.
authors.forename: Tominari
authors.initials: T
authors.lastname: Kobayashi
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan.
authors.forename: Shigeo
authors.initials: S
authors.lastname: Nagashima
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nishizawa
authors.affiliation: Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi 329-0498, Japan. Electronic address: hokamoto@jichi.ac.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okamoto
mesh_terms: D000818:Animals; D000835:Animals, Wild / Q000821:virology*; D045744:Cell Line, Tumor; D016751:Hepatitis E / Q000821:virology*; D016752:Hepatitis E virus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D051381:Rats / Q000821:virology*; D014776:Virus Cultivation; D014779:Virus Replication
keywords: Cell culture; Hepatitis E virus; Human hepatoma cells; Rattus rattus; Subgenomic RNA