HUMAN MONOCYTE MODULATION OF COAGULATION/FIBRINOLYSIS

纤维蛋白和单核细胞都存在于免疫性 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 通过产生有效的促凝血活性的炎症刺激 （PCA）。 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 单核细胞的指令。 也有一些控制过程 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），其将用于阻止 纤维蛋白溶解 单核细胞分泌的UK-1增加， 暴露于炎症刺激物。 因此， 单核细胞PCA和UK-I的局部表达是重要的， 某些病变的发病机制。 事实上， 以下列疾病为标志的闭塞性血管事件的发生率 在体外增加单核细胞PCA的条件。 然而，在这方面， 单核细胞也分泌尿激酶原（proUK）。 Pro UK衍生 来自非单核细胞来源的可能具有纤溶酶原激活剂 活动 目前尚不清楚单核细胞分泌尿激酶原是否是 受炎症刺激的影响。 之间的平衡 因此，上述活动很重要。 研究 本文所概述的建议使用我们实验室的操作方法，或 i）确定合成和 人单核细胞分泌尿激酶原受 炎症刺激物（通过定量Western印迹分析，使用 的3/H-亮氨酸，以辨别由于合成与分泌 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、促纤溶和纤溶 抑制分子（使用T细胞克隆、分离的单核细胞和 每种活性的特定测定）; iii）确定 促凝血、促纤溶和纤溶的调节 抑制剂活性，即，“平衡”如何调节？ （衍生 根据上述实验的数据）; 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，或 前酶（125/I切割的直接测量） 纤溶酶原，以避免纤溶酶激活的混杂效应， 亲英国到英国。 反应混合物的蛋白质印迹以鉴定 活性（亲）UK为1或2链。）;以及v）确定以下各项的影响 尿激酶与单核细胞的结合对其生物活性的影响 （作为纤溶酶原激活剂的效率，和对UK的耐受性， （一）。

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).