Trans-encapsidation of hepatitis C virus subgenomic replicon RNA with viral structure proteins.
A trans-packaging system for hepatitis C virus (HCV) subgenomic replicon RNAs was developed. HCV subgenomic replicon was efficiently encapsidated by the HCV structural proteins that were stably expressed in trans under the control of a mammalian promoter. Infectious HCV-like particles (HCV-LPs), established a single-round infection, were produced and released into culture medium in titers of up to 10(3) focus forming units/ml. Expression of NS2 protein with structural proteins (core, E1, E2, and p7) was shown to be critical for the infectivity of HCV-LPs. Anti-CD81 treatment decreased the number of infected cells, suggesting that HCV-LPs infected cells in a CD81-dependent manner. The packaging cell line should be useful both for the production of single-round infectious HCV-LPs to elucidate the mechanisms of HCV assembly, particle formation and infection to host cells, and for the development of HCV replicon-based vaccines.
Ishii
Koji
K
Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. kishii@nih.go.jp
Murakami
Kyoko
K
Hmwe
Su Su
SS
Zhang
Bin
B
Li
Jin
J
Shirakura
Masayuki
M
Morikawa
Kenichi
K
Suzuki
Ryosuke
R
Miyamura
Tatsuo
T
Wakita
Takaji
T
Suzuki
Tetsuro
T
D000906:Antibodies / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology; D002460:Cell Line / Q000821:virology*; D015169:Colony Count, Microbial; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D006801:Humans; D012367:RNA, Viral / Q000378:metabolism; D012093:Replicon; D060225:Tetraspanin 28; D017361:Viral Nonstructural Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics; D014771:Virion / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D019065:Virus Assembly; D014776:Virus Cultivation; D053586:Virus Internalization
D000906:Antibodies; D015703:Antigens, CD; C554798:CD81 protein, human; C097010:NS2 protein, Hepatitis C virus; D012367:RNA, Viral; D060225:Tetraspanin 28; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins