HUMAN MONOCYTE MODULATION OF COAGULATION/FIBRINOLYSIS

人类单核细胞对凝血/纤维蛋白溶解的调节

• 批准号: 3074125
• 负责人: BRADFORD S SCHWARTZ
• 金额: $ 5.29万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3074125
• 关键词: T lymphocyte; antifibrinolytic agents; fibrin; fibrinolysis; fibrinolytic agents; human tissue; monocyte; plasminogen activator; radioimmunoassay; radiotracer; tissue /cell culture; urokinase

Both fibrin and monocytes are present in lesions of immune mediated tissue damage. The process of fibrin deposition in these lesions may involve monocytes, as these cells respond to many inflammatory stimuli by elaborating potent procoagulant activity (PCA). Indeed, expression of monocyte PCA seems to be an intrinsic part of the immune response as T-lymphocytes are required for recognition of specific stimuli, with subsequent instructions to monocytes. There are also processes which control the persistence of fibrin as it is formed. Monocytes secrete an inhibitor of urokinase (UK-I) which would serve to impede fibrinolysis. Monocyte secretion of UK-1 is augmented by exposure to inflammatory stimuli. It is therefore possible that local expression of monocyte PCA and UK-I are important in the pathogenesis of certain lesions. Indeed, there is an increased incidence of occlusive vascular events in diseases marked by conditions which increase monocyte PCA in vitro. However, monocytes also secrete prourokinase (pro UK). Pro UK derived from non monocyte sources may have plasminogen activator activity. It is not known if monocyte secretion of pro UK is influenced by inflammatory stimuli. The balance between the above activities would therefore be important. The studies outlined herein propose to use methods operative in our lab or published in the literature to: i) determine whether synthesis and secretion of pro UK by human monocytes is influenced by inflammatory stimuli (assay by quantitative Western blotting, use of 3/H-Leucine to discern changes due to synthesis vs secretion of preformed pro UK) ii) determine cellular interactions and metabolic requirements of lymphocyte and monocyte populations in modulation of procoagulant, profibrinolytic, and fibrinolytic inhibitory molecules (using T-cell clones, isolated monocytes and specific assays for each activity); iii) determine relationships in regulation of procoagulant, profibrinolytic, and fibrinolytic inhibitor activities, i.e., how is the "balance" regulated? (Derived from data of the above experiments); iv) determine whether prourokinase elaborated by monocytes is fibrinolytically active or a pro-enzyme (direct measurement of cleavage of 125/I plasminogen to avoid confounding effects of plasmin activation of pro UK to UK. Western blotting of reaction mixtures to identify active (pro) UK as 1 or 2 chains.); and v) determine the effects of binding of urokinase by monocytes on its biologic activity (efficiency as a plasminogen activator, and susceptability to UK- I).

纤维蛋白和单核细胞均存在于免疫损伤中 介导的组织损伤。 纤维蛋白沉积的过程 病变可能涉及单核细胞，因为这些细胞对许多 通过阐述有效的促凝血活性来刺激炎症 （主成分分析）。 事实上，单核细胞 PCA 的表达似乎是 免疫反应的内在部分，如 T 淋巴细胞 识别特定刺激所需的，随后 对单核细胞的指令。 还有控制过程 纤维蛋白形成时的持久性。 单核细胞分泌一种 尿激酶抑制剂（UK-I），这将有助于阻碍 纤维蛋白溶解。 UK-1 的单核细胞分泌增强 暴露于炎症刺激。 因此有可能 单核细胞 PCA 和 UK-I 的局部表达在 某些病变的发病机制。 确实有增加 疾病中血管闭塞事件的发生率 体外增加单核细胞 PCA 的条件。 然而， 单核细胞还分泌尿激酶原（pro UK）。 Pro UK 衍生 非单核细胞来源可能含有纤溶酶原激活剂 活动。 目前尚不清楚单核细胞分泌的 pro UK 是否 受炎症刺激的影响。 之间的平衡 因此，上述活动非常重要。 研究 本文概述建议使用我们实验室中有效的方法或 发表在文献中的目的是： i) 确定是否合成和 人类单核细胞分泌的 pro UK 受到以下因素的影响 炎症刺激（通过定量蛋白质印迹法测定，使用 3/H-亮氨酸以辨别由于合成与分泌引起的变化 preformed pro UK) ii) 确定细胞相互作用和 淋巴细胞和单核细胞群的代谢需求 调节促凝血、促纤维蛋白溶解和纤维蛋白溶解 抑制分子（使用 T 细胞克隆、分离的单核细胞和 每项活动的具体测定）； iii) 确定关系 促凝血、促纤维蛋白溶解和纤维蛋白溶解的调节 抑制剂的活性，即“平衡”是如何调节的？ （衍生的 来自上述实验的数据）； iv) 确定是否 单核细胞产生的尿激酶原具有纤溶活性或 酶原（直接测量 125/I 的裂解 纤溶酶原以避免纤溶酶激活的混杂效应 亲英国到英国。 对反应混合物进行蛋白质印迹鉴定 活跃（专业）英国为 1 或 2 条链。）； v) 确定影响 单核细胞与尿激酶的结合对其生物活性的影响 （作为纤溶酶原激活剂的效率，以及对英国- 我）。