Study on platelet adhesion molecule to inhibit aggregation.

血小板粘附分子抑制聚集的研究。

• 批准号: 07807202
• 负责人: MIWA Masao
• 金额: $ 1.34万
• 依托单位: University of Shizuoka
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07807202/
• 关键词: platelet-activating factor; PAF; aggregation; platelet; adhesion molecule; monoclonal antibody

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

血小板粘附在血栓形成的初始阶段起着重要作用，但对决定血小板粘附是否导致血小板单层或血小板血栓形成的过程知之甚少。本研究的目的是探讨血小板聚集和去聚集的过程，特别是血小板表面粘附分子的表达机制，首先阐明血小板活化因子（PAF）聚集的血小板可被PAF拮抗剂WEB 2086去聚集，而凝血酶则不能。血小板的去聚集是基于粘附分子在细胞表面的表达，从而使细胞与细胞之间的相互作用发生解离，我们从小鼠脾细胞中获得了产生抗兔血小板抗体的克隆（MY 2 -73），该克隆对血小板的去聚集有应答，并具有不同于其他已报道的粘附分子的功能。 ...更多信息 该单克隆抗体属于IgG_<2a>k型。该单克隆抗体<51>以剂量依赖性方式抑制载铬血小板与纤维蛋白原和胶原蛋白的结合，其对刺激血小板的反应性降低，且其表位与血小板细胞表面的GMP-140和GPIIb/GPIIIa不同。用CHAPS增溶兔血小板膜蛋白，用乳过氧化物酶标记亲和柱纯化的粘附分子<125>。用<125>MY 2 -73单克隆抗体、生物素-F（ab '）_2兔抗鼠IgG（H+L）抗体、亲和素-琼脂糖免疫沉淀法，在SDS-PAGE凝胶上分离出120和110 kDa的少量~（60）I标记的粘附分子，我们正在对大量纯化后的粘附分子蛋白的性质进行进一步研究。少

项目成果

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Masashi Mizuguchi: "Neuronal and vascular pathology as a manifestation of the central nervous system toxicity of Verotoxin 2" Acta Neuropathologica. (in press). (1996)

Masashi Mizuguchi：“神经和血管病理学是 Verotoxin 2 中枢神经系统毒性的表现”Acta Neuropathologica。

J.Sugatani, Masao Miwa, Y.Komiyama, S.Ito: J.Lipid MEDIATORS and Cell Signalling. 13 (1). 73-88 (1996)

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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)

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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)

Junko Sugatani 等人：“高密度脂蛋白抑制人血管内皮细胞中血小板激活因子的合成”J.Lipid Mediators。