1995 Fiscal Year Final Research Report Summary
Biological, biochemical, and genetical analysis of cysteine proteinases of lung flukes and its application to immunodiagnosis of paragonimiasis.
| Project/Area Number |
06670276
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | Kanazawa Medical University |
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Principal Investigator |
IKEDA Teruaki Kanazawa Medical University, Medical zoology, Associate professor, 医学部, 助教授 (70064607)
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| Co-Investigator(Kenkyū-buntansha) |
ICHIKAWA Hidetaka Kanazawa Medical University, Medical zoology, Research assistant, 医学部, 助手 (40151469)
OIKAWA Yousaburou Kanazawa Medical University, Medical zoology, Research assistant, 医学部, 助手 (10139785)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Lung fluke / Paragonimiasis / Cysteine proteinase / Monoclonal antibody / ELISA / Cystatin / Recombinant antigen |
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| Research Abstract |
Results of the studies supported in part by the Grant-in-Aid are as follows :
1.Acid and neutral cysteine proteinases of Paragonimus ohirai were purified from excretory-secretory products, using various chromatography methods.
2.An IgM monoclonal antibody which reacted with the fluke cysteine proteinases was prepared from an immunized mouse. The monoclonal antibody recognized the gut epithelium and luminal contents of the parasite and was a genus-specific antibody against Paragonimus cysteine proteinases.
3.An ELISA using the partially purified cysteine proteinases gave high sensitivity and specificity for immunodiagnosis of paragonimiasis.
4.A bacterially produced fusion protein was prepared from cDMA clones encoding P.westermani cysteine proteinases. The fusion protein can detect human sera with Paragonimus infection in immunoblot analysis.
5.For detecting fluke cysteine proteinase-specific antibodies without the need for the purified proteinases, an ELISA using cystatin of a cystein proteinase inhibitor was developed.
6.The inhibition of Paragonimus cysteine proteinases by IgG antibodies of P.ohirai-infected rats was examined using a macromolecular substrate and synthetic peptid substrates. The results suggested that the inhibition of fluke cysteine proteinases by specific IgG might be due to steric hindrance occurred by IgG binding than IgG binding to an enzyme active site.
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