title: Production of infectious ferret hepatitis E virus in a human hepatocarcinoma cell line PLC/PRF/5.
abstract: A strain of ferret hepatitis E virus (HEV), sF4370, isolated from an imported ferret was used to inoculate a human hepatocarcinoma cell line, PLC/PRF/5. The virus genome and capsid protein were detected in the cell culture supernatant. Immunofluorescence microscopy indicated that the capsid protein was located in the cytoplasm. The virus particles were purified from the culture supernatant by sucrose gradient ultracentrifugation. The capsid protein with molecular mass of ∼72 kDa was detected in fractions with density of 1.150-1.162 g/cm(3), and particles of ferret HEV was associated with cell membrane. The virus recovered from the supernatant was serially passaged with PLC/PRF/5 cells and had the ability to infect ferrets by oral inoculation, indicating that the ferret HEV grown in PLC/PRF/5 was infectious. The establishment of ferret HEV cell culture system might be useful to understand the life cycle, mechanism of infection and replication of ferret HEV.
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan. Electronic address: litc@nih.go.jp.
authors.forename: Tian-Cheng
authors.initials: TC
authors.lastname: Li
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Yoshizaki
authors.affiliation: Department of Clinical Laboratory, Affiliated Hospital of Qingdao University Medical College, Jiangsu Road 16, Qingdao 266003, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors.forename: Michiyo
authors.initials: M
authors.lastname: Kataoka
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: Division of Experimental Animals Research, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Ami
authors.affiliation: Division of Experimental Animals Research, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors.forename: Suzaki
authors.initials: S
authors.lastname: Yuriko
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0781, Japan.
authors.forename: Naokazu
authors.initials: N
authors.lastname: Takeda
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000032:analysis / Q000737:chemistry; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000821:virology; D002499:Centrifugation, Density Gradient; D003470:Culture Media / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D005289:Ferrets; D016751:Hepatitis E / Q000662:veterinary / Q000821:virology; D016752:Hepatitis E virus / Q000254:growth & development* / Q000302:isolation & purification / Q000502:physiology; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D012367:RNA, Viral / Q000032:analysis; D014771:Virion / Q000302:isolation & purification
chemical_list: D036022:Capsid Proteins; D003470:Culture Media; D012367:RNA, Viral
keywords: Cell culture; Ferret HEV; Infectivity; PLC/PRF/5