| Project/Area Number |
15580285
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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 |
Clinical veterinary science
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| Research Institution | Gifu University (2004)
Obihiro University of Agriculture and Veterinary Medicine (2003) |
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Principal Investigator |
UZUKA Yuji Gifu University, Applied biological science, professor, 応用生物科学部, 教授 (30151913)
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| Co-Investigator(Kenkyū-buntansha) |
SARASHINA Takao Obihiro University of Agriculture and Veterinary medicine, School of Veterinary Medicine, professor, 畜産学部, 教授 (00154138)
OMATA Yoshitaka Obihiro University of Agriculture and Veterinary medicine, School of Veterinary Medicine, associate professor, 畜産学部, 助教授 (10132987)
TANABE Shigeyuki Obihiro University of Agriculture and Veterinary medicine, School of Veterinary Medicine, associate professor, 畜産学部, 助教授 (70292092)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | myasthenia gravis / acetylcholine receptor / canine / insect cell expression / α-subunit / fetal type of AChR / mature type of AChR / ELISA / αサブユニット / εサブユニット / コスミックII / TE671 / 無症筋無力症 / α-サブユニット |
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| Research Abstract |
Acquired myasthenia gravis (MG) is the autoimmune neuromuscular disease due to the anti-acetylcholine receptor (AChR) antibody for the acetylcholine receptor (AChR) on postsynaptic menbrane of skeletal muscle. Anti-AChR antibodies are cllasfied by blocking antibody recognized acetylcholine binding site and binding antibody recognized main immunogenic region. In this study, the gene encoding the entire canine AChR α-subunit was cloned and inserted into an Escherichia coli expression vector and an insect cells expression vector, and subsequently expressed recombinant canine AChR α subunit pretein (re-AchR α P) respectively. Western blot analyses and ELISA for serological diagnosis of acquired MG were carried out using the re-AChR α p expressed in E. coli and in insect cells severally. The re-AChR α p was able to confirm severally, but these immunoassay with these proteins was not able to differentiate between sera from antibody-positive MG dogs and sera from antibody-negative healthy dogs. Therefore, at present stage, this assay is not useful for the clinical diagnosis of acquired MG.
In additon, we investigated the evaluation of the measuring device of anti-AChR antibody for human sera (Cosmic II), which was utilized AChR a P from rhabdomyosarcoma as a antigen. 4 measured values were relatively low and 6 were high compared from conventional measuring method in 14 canine sera, which were procured from clinical acquired MG. This implies the decreased sensitivity by different species specificity and the reinforcement sensitivity by large quantities of mature type AChR of
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