title: Hepatitis E genotype 4 virus from feces of monkeys infected experimentally can be cultured in PLC/PRF/5 cells and upregulate host interferon-inducible genes.
abstract: The understanding of the interaction between hepatitis E virus (HEV) and its host cells has been impeded greatly by the absence of a cell culture system. In this study, an efficient cultivation method was developed in PLC/PRF/5 cells for HEV genotype 4 from the feces of monkeys infected experimentally. Compared to minimal essential medium (MEM), mixed Dulbecco's Modified Eagle's Medium (DMEM)/M199 improved the infection efficiency of HEV in PLC/PRF/5 cells. The incubation time and temperature were set at 6 hr and 40°C, respectively. Compared to a 100% ELISA positive ratio (EPR) of 1 × 10(6)  copies/ml HEV inoculated flasks, the ELISA positive ratio was 100%, 75%, 37.5%, and 100% for flasks inoculated with HEV incubated for 30 min under the conditions of pH 3.0, pH 11.0, 56°C and delipidation treatment, respectively. Gene expression profiles of HEV inoculated and control PLC/PRF/5 cells were assayed using a microarray. Four interferon-inducible genes, IFI27, IFI6, Mx1, and CMPK2, were up-regulated during HEV-infection. Furthermore, the replication of HEV was inhibited at 3-14 days after treatment with 500 IU/ml IFN-α2b.
authors.affiliation: Graduate School of Peking Union Medical College, Beijing, China; Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Qi
authors.forename: Tim J
authors.initials: TJ
authors.lastname: Harrison
authors.forename: Baobin
authors.initials: B
authors.lastname: Luo
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Xiuhua
authors.initials: X
authors.lastname: Li
authors.forename: Aijing
authors.initials: A
authors.lastname: Song
authors.forename: Weijin
authors.initials: W
authors.lastname: Huang
authors.forename: Youchun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D004195:Disease Models, Animal; D005243:Feces / Q000821:virology*; D020869:Gene Expression Profiling; D005838:Genotype; D000882:Haplorhini; D016751:Hepatitis E / Q000821:virology*; D016752:Hepatitis E virus / Q000235:genetics / Q000254:growth & development* / Q000276:immunology / Q000302:isolation & purification*; D054884:Host-Pathogen Interactions; D006863:Hydrogen-Ion Concentration; D007372:Interferons / Q000096:biosynthesis* / Q000235:genetics; D013696:Temperature; D015854:Up-Regulation; D014776:Virus Cultivation
chemical_list: D003470:Culture Media; D007372:Interferons
keywords: HEV; culture; interferon; microarray