2012 Fiscal Year Final Research Report
Analysis of the replication mechanism and pathology of hepatitis E virus
| Project/Area Number |
22590429
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Virology
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| Research Institution | National Institute of Infectious Diseases |
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Principal Investigator |
KOJI Ishii 国立感染症研究所, ウイルス第二部, 室長 (40280763)
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| Co-Investigator(Kenkyū-buntansha) |
TIANCHENG Li 国立感染症研究所, ウイルス第二部, 主任研究官 (90370957)
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| Project Period (FY) |
2010 – 2012
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| Keywords | ウイルス病原性 / 治療薬 / スクリーニング / 分子生物学 |
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| Research Abstract |
Hepatitis E virus (HEV) causes acute and fulminant hepatitis E in humans. Infectious cDNA clones of HEV were established by several groups, and efficient HEV propagation in cultured cells has been recently developed using PLC/PRF/5 and A549 cells. However, our PLC/PRF/5 cells showed limited permissiveness for HEV. In this study, we performed single-cell cloning of PLC/PRF/5 cells and analyzed heterogeneity by using infectious HEV clone. We constructed an infectious cDNA clone of HEV from porcine liver. Then we performed single-cell cloning of PLC/PRF/5 cells by limited dilution. To analyze the limited permissiveness of our PLC/PRF/5 cells, cloned cells were either infected with HEV or transfected with an infectious HEV RNA and the production of HEV was measured by the detection of HEV capsid protein in culture supernatant by ELISA. Ninety-eight clones were obtained after single-cell cloning of parental PLC/PRF/5 cells. The cloned PLC/PRF/5 cells exhibited various levels of HEV virus infection efficiency, and some clones were not permissive. While the replication efficiencies measured by the transfection of infectious HEV RNA differed among the cloned PLC/PRF/5 cells, these efficiencies did not correlate with infectious permissibility. We cloned several PLC/PRF/5 cells that were nonpermissive for HEV. However, HEV could replicate in these nonpermissive cell lines, suggesting that these cell lines may have some deficiencies in the attachment or entry steps of HEV infection. Investigation of host factors responsible for these steps is in progress.
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Research Products
(27 results)
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[Journal Article] Hepatitis A outbreak associated with a revolving sushi bar in Chiba, Japan: application of molecular epidemiology.2012
Author(s)
Tominaga A., Kanda T., Akiie T., Komoda H., Ito K., Abe A., Aruga A., Kaneda S., Saito M., Kiyohara T., Wakita T., Ishii K., Yokosuka O. and Sugiura N.
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Journal Title
Hepatology Research
Volume: 42
Pages: 828-834
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[Journal Article] Possible widespread presence of hepatitis A virus subgenotype IIIA in Japan: recent trend of hepatitis A causing acute liver failure.2012
Author(s)
Ishii K., Miyamura T., Kanda T., Tawada A., Sekimoto T., Wu S., Nakamoto S., Arai M., Fujiwara K., Imazeki F., Kiyohara T., Wakita T. and Yokosuka O.
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Journal Title
Hepatology Research
Volume: 42
Pages: 248-253
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[Journal Article] Long-term persistent GBV-B infection and development of a chronic and progressive hepatitis C-like disease in marmosets.2011
Author(s)
Iwasaki Y., Mori K., Ishii K., Maki N., Iijima S., Yoshida T., Okabayashi S., Katakai Y., Lee Y.J., Saito A., Funai H., Kimura N., Ageyama N., Yoshizaki S., Suzuki T., Yasutomi Y., Miyamura T., Kannnagi M. and Akari H.
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Journal Title
Frontiers in Microbiology
Volume: 2
Pages: 240
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[Journal Article] Production of infectious hepatitis C virus by using RNA polymerase I-mediated transcription.2010
Author(s)
Masaki T., Suzuki R., Saeed M., Mori K., Matsuda M., Aizaki H., Ishii K., Maki N., Miyamura T., Matsuura M., Wakita T. and Suzuki T.
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Journal Title
Journal of Virology
Volume: 84
Pages: 5824-5835
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