| Project/Area Number |
11660299
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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 |
Basic veterinary science/Basic zootechnical science
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| Research Institution | THE UNIVERSITY OF TOKYO (2000-2001)
Kagoshima University (1999) |
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Principal Investigator |
TOHYA Yukinobu Graduate school of agricultural and life sciences, Associate Professor, 農学生命科学研究科, 助教授 (20180119)
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| Co-Investigator(Kenkyū-buntansha) |
遠矢 幸伸 鹿児島大学, 農学部, 助教授 (20180119)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | calicivirus / host specificity / receptor / dog / cat / capsid / non-structural protein / genome / 中和エピトープ / モノクローナル抗体 / プロテアーゼ |
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| Research Abstract |
The final goal of this study is to find viral and cellular factors that determine the host specificity of calicivirus by the use of feline and canine caliciviruses in cell culture systems. In this study, we obtained several new findings as follows.
1. Molecular biological analysis of canine calicivirus (CaCV) : The complete nucleotide sequence of CaCV genome was determined. Two conformational neutralizing epitopes of CaCV identified by monoclonal antibodies (MAbs) were located in regions corresponding to the 5' and 3' hypervariable regions of the capsid protein of feline calicivirus (FCV).
2. In vitro host ranges of FCV and CaCV : CaCV can only replicate in cell lines of canine organ. FCV can grow well in feline cell lines and some strains of FCV can replicate in Vero cells of simian origin. However, progeny viruses were equally produced in any cell lines by transfection of the RNA genomes from FCV and CaCV, suggesting that early interaction of the caliciviruses with cells may be the major determinant for their cell tropism in vitro.
3. Virus-binding assay : Using a MAb produced in section 1., a virus binding assay by flow cytemetry was developed for CaCV. The analysis indicated that CaCV binds efficiently to the permissive cells, but does not bind to non-permissive cells.
4. A MAb against MDCK cells of canine origin that blocks CaCV infection was produced and characterized. The MAb bound to the permissive cells but not to the non-permissive cells for CaCV and inhibited the binding of CaCV to the permissive cells, indicating that the MAb recognizes the receptor for CaCV. By immunoprecipitation, the MAb was shown to react with two proteins with molecular masses of 76 and 65 kDa.
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