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Study on platelet adhesion molecule to inhibit aggregation.

Research Project

Project/Area Number07807202
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionUniversity of Shizuoka

Principal Investigator

MIWA Masao  School of Pharmaceutical Sciences, University of Shizuoka, Professor, 薬学部, 教授 (10046287)

Project Period (FY) 1995 – 1996
Project Status Completed(Fiscal Year 1996)
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)
Keywordsplatelet-activating factor / PAF / aggregation / platelet / adhesion molecule / monoclonal antibody
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

Report

(3results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report

Research Products

(7results)

All Other

All Publications

  • [Publications] Junko Sugatani et al.: "High-density lipiprotein inhibits the synthesis of platelet-activating factor in human vascular endothelial cells" J.Lipid.Mediators. 13. 73-88 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Masashi Mizuguchi et al: "Neuronal and vascular pathology produced by verocytotoxin 2 in the rabbit central nervous system" Acta Neuropathol. 91. 254-262 (1966)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] J.Sugatani, Masao Miwa, Y.Komiyama, S.Ito: J.Lipid MEDIATORS and Cell Signalling. 13 (1). 73-88 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Junko Sugatani et al.: "High-density lipiprotein inhibits the synthesis of platelet-activating factor in human vascular endothelial cells" J. Lipid Mediators. 13. 73-88 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Masashi Mizuguchi et al: "Neuronal and vascular pathology produced by verocytotoxin 2 in the rabbit central nervous system" Acta Neuropathol. 91. 254-262 (1966)

    • Related Report
      1996 Annual Research Report
  • [Publications] Junko Sugatani: "High density lipoprotein inhibits the synthesis of platelet-activating factor in human endothelial cells" J.Lipid mediators Cell Signalling. 73. 73-88 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Masashi Mizuguchi: "Neuronal and vascular pathology as a manifestation of the central nervous system toxicity of Verotoxin 2" Acta Neuropathologica. (in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published : 1995-04-01   Modified : 2016-04-21  

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