HUMAN MONOCYTE MODULATION OF COAGULATION/FIBRINOLYSIS

纤维蛋白和单核细胞都存在于免疫介导的组织损伤中 损害 这些病变中纤维蛋白沉积的过程可能涉及 单核细胞，因为这些细胞对许多炎症刺激作出反应， 产生有效的促凝血活性（PCA）。 事实上， 单核细胞PCA似乎是免疫应答的固有部分， T淋巴细胞是识别特定刺激所必需的， 随后的指令给单核细胞。 也有一些过程， 控制纤维蛋白形成时的持久性。 单核细胞分泌 尿激酶抑制剂（UK-I），其可用于阻止纤维蛋白溶解。 暴露于炎性介质可增加单核细胞分泌UK-I 刺激。 因此，单核细胞PCA的局部表达可能是 和UK-I在某些病变的发病机制中是重要的。 的确， 疾病中闭塞性血管事件的发生率增加 以增加体外单核细胞PCA的条件为标志。 然而，在这方面， 单核细胞也分泌尿激酶原（proUK）。 Pro UK源自非 单核细胞来源可以具有纤溶酶原激活剂活性。 目前还不知道 如果单核细胞分泌尿激酶原受炎症刺激的影响。 的 因此，上述活动之间的平衡很重要。 的 本文概述的研究建议使用我们实验室中可操作的方法， i）确定合成和 人单核细胞分泌尿激酶原受炎症因子的影响 刺激（通过定量Western印迹分析，使用3 H-亮氨酸， 辨别由于预形成的尿激酶原的合成与分泌引起的变化）; ii） 确定淋巴细胞的细胞相互作用和代谢要求 和单核细胞群调节促凝血，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 单核细胞和每种活性的特异性测定）; iii）确定 调节促凝血、促纤溶和 纤维蛋白溶解抑制剂活性，即，“平衡”如何调节？ （源自上述实验的数据）; iv）确定是否 由单核细胞加工的尿激酶原具有纤维蛋白溶解活性， 前酶（直接测量125 I纤溶酶原的裂解，以避免 纤溶酶激活pro UK对UK的混淆效应。 西方 印迹反应混合物以鉴定活性（原）UK为1或2 链）;和v）测定尿激酶与单核细胞结合的影响 其生物活性（作为纤溶酶原激活剂的效率，以及 对UK-I的敏感性）。

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-I 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 3H-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 125I 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 susceptibility to UK-I).