| Project/Area Number |
11557025
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Virology
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| Research Institution | The Tokyo Metropolitan Institute of Medical Science |
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Principal Investigator |
KOHARA Michinori The Tokyo Metropolitan Institute of Medical Science, Head, 東京都臨床医学総合研究所, 研究員 (10250218)
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| Co-Investigator(Kenkyū-buntansha) |
FUJITA Takasi The Tokyo Metropolitan Institute of Medical Science, Head, 医科学研究所, 研究員 (10156870)
YONEKAWA Hiromichi The Tokyo Metropolitan Institute of Medical Science, Vice Director, 医科学研究所, 研究員 (30142110)
KOHARA Kyoko The Tokyo Metropolitan Institute of Medical Science, Research Scientist, 医科学研究所, 講師 (20225478)
TSUCHIYA Masayuki Chugai, Research Scientist, 創薬資源研究所, 研究員
TOYODA Tetsuya Kurume University School of Medicine, Professor, 医学部, 教授 (00197972)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 2000: ¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1999: ¥6,800,000 (Direct Cost: ¥6,800,000)
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| Keywords | HCV / infectious cDNA / pathogenesis / replication / インターフェロン / シグナル伝達 / HCV cDNAクローン |
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| Research Abstract |
[Purpose] Previous studies of transmission of hepatitis C virus (HCV) by intrahepatic inoculation with synthetic RNA have been reported. However, rescue of viable HCV particle from these chimpanzees or in vitro culture system had not be successful, so far. In this study, we established reverse genetical system of HCV to characterize genome function. [Methods] We constructed the entire HCV cDNA clone from chronic hepatitis patient plasma and expressed exact mRNA of HCV genome using double ribozyme trimming system. The RNA transcripts from this clone have been transfected into IMY-N9 cells, which was the fused cell line of HepG2 and human primaly hepatocyte. [Results and conclusions] Transfection of the synthetic HCV RNA and cDNA into IMY-N9 cells produced infectios HCV, however in HepG2 cells, they could not support HCV replication. The particle with density of 1.1 g/ml in the culture supernatant was visualized by immunoelectron microscopic study. This particle was considered to be the mature particle. This culture supernatant contained HCV particles even after the DNase and RNase treatment and could re-infect to the naive IMY cells and could cause replication. These results indicated that this cDNA clone rescued the infectious HCV particle and has opened the possibility to the reverse genetical approach of HCV.
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