Adenovirus-mediated gene transfer of interferon alpha inhibits hepatitis C virus replication in hepatocytes.
Recently we reported that on-site interferon (IFN)-alpha production in the liver using an adenovirus vector can achieve a substantial confinement of IFN-alpha in the target organ and can improve liver fibrosis in a rat liver cirrhosis model. However, the major therapeutic effect of IFN for hepatitis C virus (HCV)-associated liver diseases is its antiviral effect on HCV. As a prelude to the in vivo HCV infection experiment using a primate animal model, here we examined the antiviral effect of IFN-alpha gene transfer into HCV-positive hepatocytes in vitro. The non-neoplastic human hepatocyte cell line PH5CH8 was inoculated with HCV-positive serum. Successful in vitro HCV replication and thus the validity of this model was confirmed by a strong selection for HCV variants determined by sequence analysis of the hypervariable region and an increase of HCV RNA estimated by real time TaqMan RT-PCR. One day after the inoculation of HCV, PH5CH8 cells were infected with adenoviral vectors encoding human IFN-alpha cDNA. HCV completely disappeared 9 days after the adenoviral infection, which is linked to the increase of 2('),5(')-oligoadenylate synthetase activity, suggesting that IFN-alpha produced by gene transfer effectively inhibits HCV replication in hepatocytes. This study supports the development of IFN-alpha gene therapy for HCV-associated liver diseases.
Suzuki
Koichi
K
Genetics Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
Aoki
Kazunori
K
Ohnami
Shumpei
S
Yoshida
Kimiko
K
Kazui
Teruhisa
T
Kato
Nobuyuki
N
Inoue
Kazuaki
K
Kohara
Michinori
M
Yoshida
Teruhiko
T
D015088:2',5'-Oligoadenylate Synthetase / Q000378:metabolism; D000256:Adenoviridae / Q000235:genetics*; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000378:metabolism; D002460:Cell Line; D005822:Genetic Vectors; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000502:physiology*; D022781:Hepatocytes / Q000201:enzymology / Q000821:virology*; D006801:Humans; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000032:analysis; D014162:Transfection; D014779:Virus Replication
D000998:Antiviral Agents; D016898:Interferon-alpha; D012367:RNA, Viral; D015088:2',5'-Oligoadenylate Synthetase