CELL CELL INTERACTIONS IN THROMBOSIS

血栓形成中的细胞相互作用

• 批准号: 2223388
• 负责人: Aaron Jacob Marcus
• 金额: $ 18.42万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2223388
• 关键词: SDS polyacrylamide gel electrophoresis; adenosine; adenosine diphosphate; adenosine monophosphate; antithrombins; arachidonate; aspirin; atherosclerosis; blood chemistry; cell cell interaction; cytokine; eicosanoid metabolism; endopeptidases; enzyme mechanism; erythrocytes; high performance liquid chromatography; human tissue; monoclonal antibody; neutrophil; nitric oxide; nucleotidases; nucleotides; platelet activation; platelet aggregation; platelets; prostacyclins; protease inhibitor; prothrombin; radioimmunoassay; thin layer chromatography; thrombosis; tissue /cell culture; vascular endothelium; SDS polyacrylamide gel electrophoresis; adenosine; adenosine diphosphate; adenosine monophosphate; antithrombins; arachidonate; aspirin; atherosclerosis; blood chemistry; cell cell interaction; cytokine; eicosanoid metabolism; endopeptidases; enzyme mechanism; erythrocytes; high performance liquid chromatography; human tissue; monoclonal antibody; neutrophil; nitric oxide; nucleotidases; nucleotides; platelet activation; platelet aggregation; platelets; prostacyclins; protease inhibitor; prothrombin; radioimmunoassay; thin layer chromatography; thrombosis; tissue /cell culture; vascular endothelium

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.

广泛的长期目标是研究 血栓形成，现在被确定为一组 多细胞事件。 接触和不同程度的病理 血管和血细胞之间的相互作用控制发育和 闭塞性疾病的可逆性。 几个体外模型系统 本文描述了这些细胞细胞的组件 互动。 这包括不同的跨细胞代谢 细胞，导致具有新型生物学活性的代谢产物。 这 提出的研究将定义生化和功能作用 介体和内在细胞活动，以增强或预防 血小板事件。 具体而言，血小板与：i）的相互作用 人内皮细胞（EC），II）中性粒细胞和III）红细胞 将被调查。 在i）初步研究表明EC 将扩大ecto-ADPase（S）块血小板反应性。 EC ecto-Adpase（S）将被隔离和表征。单克隆抗体 将开发并确定ADPase的调节。 第二个 i）的成分涉及内皮衍生的放松因子（EDRF/no）， 一种抑制血细胞和血管反应性的自闭症。 EC和中性粒细胞的EDRF/NO形成。 EC/NO 合成酶系统将被隔离。 3个主的效果 抗血栓性防御机制（EDRF/NO，类固醇和ADPases） 将是区分和相关的。 在ii）抑制血小板 中性粒细胞的激活和募集将分为EDRF/NO 相关和无关的组件。 中性粒细胞激活的影响和 关于抗血栓性活动的产生将是 决定。 中性粒细胞抑制性活动的稳定性允许 用于本地化，隔离以及可能的转移。 在III中） 血小板激活的代谢促进的基础机制和 红细胞招募将阐明，包括影响 去除核苷酸，蛋白酶抑制和阿司匹林治疗。 已建立的实验方法包括：单独的测量 血小板激活和募集，组织培养，TLC，HPLC，GC/MS， 聚合和放射免疫测定法。