CELL-CELL INTERACTIONS IN THROMBOSIS

广泛的长期目标是研究 血栓形成，这是现在成立的一个综合组， 多细胞事件 接触和不同程度的病理性 血管和血细胞之间的相互作用决定着发育， 闭塞性疾病的可逆性。 几种体外模型系统 来研究这些细胞-细胞 交互. 这包括不同细胞之间的跨细胞代谢， 细胞，产生具有新的生物活性的代谢物。 的 拟议的研究将确定生物化学和功能作用， 介质和内在细胞活动，增强或防止 血栓形成事件。 具体而言，血小板与以下物质的相互作用： 人内皮细胞（EC），II）嗜中性粒细胞，和III）红细胞 将被调查。 （一）初步研究表明， 将延长外腺苷酸阻断血小板反应性。 EC 将分离和表征外接ADRs。单克隆抗体 将被开发并确定ADD 1000的监管。 第二 I）的组分涉及内皮源性舒张因子（EDRF/NO）， 一种抑制血细胞和血管反应的自体素。 将表征EC和中性粒细胞形成EDRF/NO。EC/NO 合成酶系统将被分离。 3主要影响 抗血栓防御机制（EDRF/NO、类花生酸和ADP酶） 将被区分和关联。 II）血小板抑制 中性粒细胞的激活和募集将分为EDRF/NO 相关和不相关的成分。 中性粒细胞活化和 对产生抗血栓活性的预充将是 测定 中性粒细胞抑制活性的稳定性允许 进行定位隔离可能还有转移 （III） 代谢促进血小板活化的潜在机制， 将阐明红细胞的募集，包括 核苷酸去除、蛋白酶抑制和阿司匹林治疗。 已建立的实验方法包括： 血小板活化和募集，组织培养，TLC，HPLC，GC/MS， 聚集测定法和放射免疫测定法。

The broad, long-term objectives are to study the pathogenesis of thrombosis, which is now established as an integrated group of multicellular events. Contact and varying degrees of pathologic interactions between vascular and blood cells govern development and reversibility of occlusive disorders. Several in vitro model systems are described herein to study components of these cell-cell interactions. This includes transcellular metabolism between different cells, resulting in metabolites with novel biological activities. The research proposed will define biochemical and functional roles of mediators and intrinsic cell activities which enhance or prevent thrombotic events. Specifically, interactions of platelets with: I) human endothelial cells (EC), II) neutrophils, and III) erythrocytes will be investigated. In I) preliminary studies indicating that EC ecto-ADPase(s) block platelet reactivity will be extended. EC ecto-ADPase(s) will be isolated and characterized. Monoclonal antibodies will be developed and regulation of ADPase(s) determined. The second component of I) involves endothelium-derived relaxing factor (EDRF/NO), an autacoid which inhibits blood cell and vascular responsiveness. Formation of EDRF/NO by EC and neutrophils will be characterized. EC/NO synthase system(s) will be isolated. Effects of the 3 main antithrombotic defense mechanisms (EDRF/NO, eicosanoids, and ADPases) will be differentiated and correlated. In II) inhibition of platelet activation and recruitment by neutrophils will be separated into EDRF/NO related and unrelated components. Effects of neutrophil activation and priming on the generation of antithrombotic activities will be determined. Stability of the neutrophil inhibitory activitie(s) allow for localization, isolation, and, possibly, transfer. In III) mechanisms underlying the metabolic promotion of platelet activation and recruitment by erythrocytes will be elucidated, including the influence of nucleotide removal, protease inhibition, and aspirin treatment. Established experimental methods include: Separate measurement of platelet activation and recruitment, tissue culture, TLC, HPLC, GC/MS, aggregometry and radioimmunoassay.