Study on platelet adhesion molecule to inhibit aggregation.
| Project/Area Number |
07807202
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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 |
Biological pharmacy
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| Research Institution | University of Shizuoka |
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Principal Investigator |
MIWA Masao School of Pharmaceutical Sciences, University of Shizuoka, Professor, 薬学部, 教授 (10046287)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | platelet-activating factor / PAF / aggregation / platelet / adhesion molecule / monoclonal antibody |
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| Research Abstract |
Platelets play an important role in the initial stage of thrombosis, but the processes that determine whether platelet adhesion results in a platelet monolayr or platelet thrombus formation is poorly understood. The purpose of the present study was to examine the events involved in platelet aggregation and desaggregation, especially the mechanism to express the adhesion molecule to platelet cell surface that responds to the desaggregation.
First, in this study, we elucidated that platelets aggregated by platelet-activating factor (PAF), known as phospholipid mediator, are deaggregated by PAF antagonist WEB2086 but not those by thrombin. The platelet desaggregation was based on the expression of adhesion molecule to cell surface which dissociate cell-cell interaction.
Furthermore, we obtained the clone (MY2-73) to produce anti-rabbit platelet antibody from mouse splenocyte, which responds to platelet desaggregation and functions as adhesion molecule different from others already reported.
… More
The monoclonal antibody was classified into IgG_<2a> type k. This monoclonal antibody prevented ^<51>Cr-loaded platelet to bind to fibrinogen and collagen in a dose-dependent manner, of which the reactivity to stimulated platelets decreased and the epitope is different from GMP-140 and GPIIb/GPIIIa on platelet cell surface.
In order to purify the adhesion molecule recognized by the monoclonal antibody, we prepared the affinity column bound to the monoclonal antibody purified from mouse ascites, and solubillized rabbit platelet membrane proteins with CHAPS.The adhesion molecules purified by the affinity column were labelled with ^<125>I using lactoperoxidase. ^<125>I-labelled adhesion molecules immunoprecipitated with MY2-73 monoclonal antibody, biotin-F (ab')_2 rabbit anti-mouse IgG (H+L) antibody, avidin-agarose were separated on SDS-PAGE gel, of which the molecular weight were 120 and 110 kDa and the amounts were very small.
We are in progress to further study the properties of adhesion molecule proteins after purification of high amounts these molecules. Less
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Report
(3 results)
Research Products
(7 results)
