title: Replication-competent recombinant vesicular stomatitis virus encoding hepatitis C virus envelope proteins.
abstract: Although in vitro replication of the hepatitis C virus (HCV) JFH1 clone of genotype 2a (HCVcc) has been developed, a robust cell culture system for the 1a and 1b genotypes, which are the most prevalent viruses in the world and resistant to interferon therapy, has not yet been established. As a surrogate virus system, pseudotype viruses transiently bearing HCV envelope proteins based on the vesicular stomatitis virus (VSV) and retrovirus have been developed. Here, we have developed a replication-competent recombinant VSV with a genome encoding unmodified HCV E1 and E2 proteins in place of the VSV envelope protein (HCVrv) in human cell lines. HCVrv and a pseudotype VSV bearing the unmodified HCV envelope proteins (HCVpv) generated in 293T or Huh7 cells exhibited high infectivity in Huh7 cells. Generation of infectious HCVrv was limited in some cell lines examined. Furthermore, HCVrv but not HCVpv was able to propagate and form foci in Huh7 cells. The infection of Huh7 cells with HCVpv and HCVrv was neutralized by anti-hCD81 and anti-E2 antibodies and by sera from chronic HCV patients. The infectivity of HCVrv was inhibited by an endoplasmic reticulum alpha-glucosidase inhibitor, N-(n-nonyl) deoxynojirimycin (Nn-DNJ), but not by a Golgi mannosidase inhibitor, deoxymannojirimycin. Focus formation of HCVrv in Huh7 cells was impaired by Nn-DNJ treatment. These results indicate that the HCVrv developed in this study can be used to study HCV envelope proteins with respect to not only the biological functions in the entry process but also their maturation step.
authors.affiliation: Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Tani
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Komoda
authors.forename: Eiko
authors.initials: E
authors.lastname: Matsuo
authors.forename: Kensuke
authors.initials: K
authors.lastname: Suzuki
authors.forename: Itsuki
authors.initials: I
authors.lastname: Hamamoto
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Yamashita
authors.forename: Kohji
authors.initials: K
authors.lastname: Moriishi
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Fujiyama
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kanto
authors.forename: Norio
authors.initials: N
authors.lastname: Hayashi
authors.forename: Ania
authors.initials: A
authors.lastname: Owsianka
authors.forename: Arvind H
authors.initials: AH
authors.lastname: Patel
authors.forename: Michael A
authors.initials: MA
authors.lastname: Whitt
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Matsuura
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D065089:Glycoside Hydrolase Inhibitors; D016174:Hepacivirus / Q000187:drug effects / Q000502:physiology*; D018937:Hepatitis C Antibodies / Q000494:pharmacology; D006801:Humans; D018942:Macrolides / Q000494:pharmacology; D008954:Models, Biological; D060225:Tetraspanin 28; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication
chemical_list: D015703:Antigens, CD; C554798:CD81 protein, human; C087035:E1 protein, Hepatitis C virus; D004791:Enzyme Inhibitors; D065089:Glycoside Hydrolase Inhibitors; D018937:Hepatitis C Antibodies; D018942:Macrolides; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus; C040929:bafilomycin A1