Trans-complemented hepatitis C virus particles as a versatile tool for study of virus assembly and infection.
In this study, we compared the entry processes of trans-complemented hepatitis C virus particles (HCVtcp), cell culture-produced HCV (HCVcc) and HCV pseudoparticles (HCVpp). Anti-CD81 antibody reduced the entry of HCVtcp and HCVcc to almost background levels, and that of HCVpp by approximately 50%. Apolipoprotein E-dependent infection was observed with HCVtcp and HCVcc, but not with HCVpp, suggesting that the HCVtcp system is more relevant as a model of HCV infection than HCVpp. We improved the productivity of HCVtcp by introducing adapted mutations and by deleting sequences not required for replication from the subgenomic replicon construct. Furthermore, blind passage of the HCVtcp in packaging cells resulted in a novel mutation in the NS3 region, N1586D, which contributed to assembly of infectious virus. These results demonstrate that our plasmid-based system for efficient production of HCVtcp is beneficial for studying HCV life cycles, particularly in viral assembly and infection.
Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. ryosuke@nih.go.jp
Ryosuke
R
Suzuki
Kenji
K
Saito
Takanobu
T
Kato
Masayuki
M
Shirakura
Daisuke
D
Akazawa
Koji
K
Ishii
Hideki
H
Aizaki
Yumi
Y
Kanegae
Yoshiharu
Y
Matsuura
Izumu
I
Saito
Takaji
T
Wakita
Tetsuro
T
Suzuki
D005816:Genetic Complementation Test; D016174:Hepacivirus / Q000472:pathogenicity* / Q000502:physiology; D006801:Humans; D014773:Virology / Q000379:methods*; D019065:Virus Assembly; D014776:Virus Cultivation / Q000379:methods; D053586:Virus Internalization