HUMAN MONOCYTE MODULATION OF COAGULATION/FIBRINOLYSIS

免疫损伤中同时存在纤维蛋白和单核细胞。 介导的组织损伤。纤维蛋白在这些组织中沉积的过程 损伤可能涉及单核细胞，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 (PCA)。事实上，单核细胞PCA的表达似乎是一种 作为免疫反应的固有部分，T淋巴细胞是 识别特定刺激所需的，随后 单核细胞的使用说明。还有一些进程可以控制 纤维蛋白在形成时的持久性。单核细胞分泌An 尿激酶抑制物(UK-I)，这将阻碍 纤溶作用。单核细胞对UK-1的分泌增加 暴露在炎性刺激下。因此，有可能 单核细胞Pca和UK-I的局部表达在骨质疏松症中起重要作用 某些损伤的发病机制。事实上，有一个增加的 闭塞血管事件在以下疾病中的发生率 体外增加单核细胞PCA的条件。然而， 单核细胞也分泌尿激酶原(尿激酶原)。亲英国派生的 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Pro UK的单核细胞分泌是否 受炎症刺激的影响。两者之间的平衡 因此，上述活动将是重要的。这些研究 在此概述的建议使用我们实验室中可操作的方法或 发表在文献中，以：i)确定合成和 人单核细胞分泌尿蛋白原受以下因素影响 炎性刺激(定量Western blotting分析，使用 3/H-亮氨酸的合成和分泌的变化 预制PRO UK)ii)确定细胞相互作用和 淋巴细胞和单核细胞群体的代谢需要 在调节促凝剂、纤溶原和纤溶方面 抑制分子(使用T细胞克隆、分离的单核细胞和 每项活动的具体分析)；三)确定 促凝剂、纤溶原和纤溶的调节 抑制活性，即“平衡”是如何调节的？(派生的 根据上述实验的数据)；四)确定 由单核细胞形成的尿激酶原具有纤溶活性或 A酶原(直接测量125/I的裂解 纤溶酶原可避免纤溶酶激活的混杂效应 亲英国对英国。反应混合物的Western blotting鉴定 活性(PRO)UK作为1或2个链)；以及v)确定 单核细胞与尿激酶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UK- i)。

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).