Serum-derived hepatitis C virus infectivity in interferon regulatory factor-7-suppressed human primary hepatocytes.
OBJECTIVE
The development of an efficient in vitro infection system for HCV is important in order to develop new anti-HCV strategy. Only Huh7 hepatocyte cell lines were shown to be infected with JFH-1 fulminant HCV-2a strain and its chimeras. Here we aimed to establish a primary hepatocyte cell line that could be infected by HCV particles from patients' sera.


METHODS
We transduced primary human hepatocytes with human telomerase reverse transcriptase together with human papilloma virus 18/E6E7 (HPV18/E6E7) genes or simian virus large T gene (SV40 T) to immortalize cells. We also established the HPV18/E6E7-immortalized hepatocytes in which interferon regulatory factor-7 was inactivated. Finally we analyzed HCV infectivity in these cells.


RESULTS
Even after prolonged culture HPV18/E6E7-immortalized hepatocytes exhibited hepatocyte functions and marker expression and were more prone to HCV infection than SV40 T-immortalized hepatocytes. The susceptibility of HPV18/E6E7-immortalized hepatocytes to HCV infection was further improved, in particular, by impairing signaling through interferon regulatory factor-7.


CONCLUSIONS
HPV18/E6E7-immortalized hepatocytes are useful for the analysis of HCV infection, anti-HCV innate immune response, and screening of antiviral agents with a variety of HCV strains.
Aly
Hussein H
HH
Graduate School of Medicine, Department of Transplant Surgery, Kyoto University Hospital, Kyoto, Japan.
Watashi
Koichi
K
Hijikata
Makoto
M
Kaneko
Hiroyasu
H
Takada
Yasutugu
Y
Egawa
Hiroto
H
Uemoto
Shinji
S
Shimotohno
Kunitada
K
D015703:Antigens, CD / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D005838:Genotype; D006367:HeLa Cells; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity / Q000502:physiology*; D006526:Hepatitis C / Q000821:virology; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology / Q000821:virology*; D052161:Human papillomavirus 18 / Q000235:genetics; D006801:Humans; D050839:Interferon Regulatory Factor-7 / Q000037:antagonists & inhibitors* / Q000235:genetics; D007370:Interferon Type I / Q000494:pharmacology; D009856:Oncogene Proteins, Viral / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D011994:Recombinant Proteins; D019098:Telomerase / Q000235:genetics; D060225:Tetraspanin 28; D014162:Transfection
D015703:Antigens, CD; C554798:CD81 protein, human; D017931:DNA Primers; D004268:DNA-Binding Proteins; C052603:E6 protein, Human papillomavirus type 18; C052633:E7 protein, Human papillomavirus type 18; D050839:Interferon Regulatory Factor-7; D007370:Interferon Type I; D009856:Oncogene Proteins, Viral; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D060225:Tetraspanin 28; C509186:TERT protein, human; D019098:Telomerase