title: Development of mouse hepatocyte lines permissive for hepatitis C virus (HCV).
abstract: The lack of a suitable small animal model for the analysis of hepatitis C virus (HCV) infection has hampered elucidation of the HCV life cycle and the development of both protective and therapeutic strategies against HCV infection. Human and mouse harbor a comparable system for antiviral type I interferon (IFN) induction and amplification, which regulates viral infection and replication. Using hepatocytes from knockout (ko) mice, we determined the critical step of the IFN-inducing/amplification pathways regulating HCV replication in mouse. The results infer that interferon-beta promoter stimulator (IPS-1) or interferon A receptor (IFNAR) were a crucial barrier to HCV replication in mouse hepatocytes. Although both IFNARko and IPS-1ko hepatocytes showed a reduced induction of type I interferons in response to viral infection, only IPS-1-/- cells circumvented cell death from HCV cytopathic effect and significantly improved J6JFH1 replication, suggesting IPS-1 to be a key player regulating HCV replication in mouse hepatocytes. We then established mouse hepatocyte lines lacking IPS-1 or IFNAR through immortalization with SV40T antigen. Expression of human (h)CD81 on these hepatocyte lines rendered both lines HCVcc-permissive. We also found that the chimeric J6JFH1 construct, having the structure region from J6 isolate enhanced HCV replication in mouse hepatocytes rather than the full length original JFH1 construct, a new finding that suggests the possible role of the HCV structural region in HCV replication. This is the first report on the entry and replication of HCV infectious particles in mouse hepatocytes. These mouse hepatocyte lines will facilitate establishing a mouse HCV infection model with multifarious applications.
authors: Department of Microbiology and Immunology, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, Japan.
authors: Hussein Hassan
authors: HH
authors: Aly
authors: Hiroyuki
authors: H
authors: Oshiumi
authors: Hiroaki
authors: H
authors: Shime
authors: Misako
authors: M
authors: Matsumoto
authors: Taka
authors: T
authors: Wakita
authors: Kunitada
authors: K
authors: Shimotohno
authors: Tsukasa
authors: T
authors: Seya
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D003588:Cytopathogenic Effect, Viral; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D016679:Genome, Viral / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D006526:Hepatitis C / Q000378:metabolism / Q000821:virology; D022781:Hepatocytes / Q000166:cytology* / Q000821:virology*; D006801:Humans; D007372:Interferons / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D053633:Receptor, Interferon alpha-beta / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D060225:Tetraspanin 28; D014162:Transfection; D014779:Virus Replication / Q000502:physiology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D015703:Antigens, CD; D007372:Interferons; D053633:Receptor, Interferon alpha-beta; D060225:Tetraspanin 28; C554798:CD81 protein, human; C554799:Cd81 protein, mouse; C512612:MAVS protein, mouse; C508032:Ifnar1 protein, mouse