Cell culture and in vivo analyses of cytopathic hepatitis C virus mutants.
HCV-JFH1 yields subclones that develop cytopathic plaques (Sekine-Osajima Y, et al., Virology 2008; 371:71). Here, we investigated viral amino acid substitutions in cytopathic mutant HCV-JFH1 clones and their characteristics in vitro and in vivo. The mutant viruses with individual C2441S, P2938S or R2985P signature substitutions, and with all three substitutions, showed significantly higher intracellular replication efficiencies and greater cytopathic effects than the parental JFH1 in vitro. The mutant HCV-inoculated mice showed significantly higher serum HCV RNA and higher level of expression of ER stress-related proteins in early period of infection. At 8 weeks post inoculation, these signature mutations had reverted to the wild type sequences. HCV-induced cytopathogenicity is associated with the level of intracellular viral replication and is determined by certain amino acid substitutions in HCV-NS5A and NS5B regions. The cytopathic HCV clones exhibit high replication competence in vivo but may be eliminated during the early stages of infection.
Mishima
Kako
K
Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8519, Japan.
Sakamoto
Naoya
N
Sekine-Osajima
Yuko
Y
Nakagawa
Mina
M
Itsui
Yasuhiro
Y
Azuma
Seishin
S
Kakinuma
Sei
S
Kiyohashi
Kei
K
Kitazume
Akiko
A
Tsuchiya
Kiichiro
K
Imamura
Michio
M
Hiraga
Nobuhiko
N
Chayama
Kazuaki
K
Wakita
Takaji
T
Watanabe
Mamoru
M
D019943:Amino Acid Substitution; D000818:Animals; D045744:Cell Line, Tumor; D003588:Cytopathogenic Effect, Viral; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity* / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D051379:Mice; D009154:Mutation; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics*; D010948:Viral Plaque Assay / Q000379:methods; D014779:Virus Replication; D012324:RNA-Dependent RNA Polymerase
D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase