| Project/Area Number |
11470515
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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 | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
MATSUNO Kazuhiko Hokkaido Univ., College of Medical Technology, Professor, 医療技術短期大学部, 教授 (70102332)
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| Co-Investigator(Kenkyū-buntansha) |
CHIBA Hitoshi Hokkaido Univ., Medical hospital School of Med., Lec., 医学部・附属病院, 講師 (70197622)
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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 |
¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1999: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | platelets / flow cytometry / CD62P / CD63 / PAC-1 / CD36 / oxidized LDL / 活性化 / β-トロンボグロブリン |
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| Research Abstract |
Platelets play key roles in the pathophysiology of thrombosis. It is important to evaluate platelet activation in vivo in order to predict the possibility of thrombosis. The purpose of this study is to establish optimized methods to monitor the activation status of circulating platelets in patients. We investigated the sensitivity of various monoclonal antibodies (MoAbs) against activation-dependent antigen (CD62P, CD63, PAC-1, CD107a, CD107b, SPAN12, WADE15) firstly. MoAbs against CD62P, CD63, PAC-1 was the most sensitive for the detection of platelet activation in vitro. Secondarily we studied the extent of ex vivo platelet activation after blood collection. A significant reduction in basal CD62P and CD63 expression was obtained when blood was fixed immediately after blood collection. However, it was necessary to incubate whole blood anticogulated with EDTA or Na citrate with anti-PAC-1 MoAb before fixation with paraformaldehyde concerning PAC-1 assay. We compared the difference of 4 methods to measure the activation of circulating platelets. No significant difference was observed in 4 methods when whole blood was fixed with paraformaldehyde right after blood collection. But it is necessary to collect blood with anticoagulant mixture (EDTA + Na citrate + adenosine + theophyllin) when whole blood is left as it is for 0.5〜1 hour. The expression of CD62P, CD63 and PAC-1 on unstimulated platelets was significantly higher in patients with dialysis than that in normal controls. ADP-stimulated expression of CD62P and CD63 on platelets was significantly higher in patients with dialysis and nephritic syndrome than that in normal controls. Oxidized-LDL-stimulated expression of CD62P on platelets was significantly lower in CD36 negative population than in CD36 positive population.
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