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病变中。 损坏。这些病变中纤维蛋白沉积的过程可能涉及 单核细胞，因为这些细胞通过 阐述了有效的促凝活性(PCA)。事实上，表达 单核细胞PCA似乎是免疫反应的固有部分，因为 T淋巴细胞是识别特定刺激所必需的，具有 单核细胞的后续指令。也有一些进程 在纤维蛋白形成时控制其持久性。单核细胞分泌An 尿激酶抑制物(UK-I)，可用于阻止纤溶。 炎症性环境中单核细胞分泌UK-I的增加 刺激物。因此，单核细胞PCA的局部表达是可能的 和UK-I在某些病变的发病机制中起重要作用。的确， 在疾病中，闭塞血管事件的发生率增加。 以在体外增加单核细胞PCA的条件为特征的。然而， 单核细胞也分泌尿激酶原(尿激酶原)。PRO UK源自非 单核细胞来源可能具有纤溶酶原激活剂活性。目前尚不清楚 单核细胞分泌尿蛋白原是否受炎症刺激的影响。这个 因此，上述活动之间的平衡将是重要的。这个 这里概述的研究建议使用我们实验室操作的方法或 发表在文献中，以：i)确定合成和 炎症对人单核细胞分泌尿蛋白原的影响 刺激物(定量Western blotting分析，使用~3H-亮氨酸 识别合成与分泌预制尿蛋白的变化)；ii) 确定淋巴细胞的细胞相互作用和代谢需求 以及单核细胞群在调节促凝血、纤溶原、 和纤溶抑制分子(使用T细胞克隆，分离 单核细胞和每种活动的特定分析)；三)确定 促凝血剂、纤溶原和血管紧张素转换酶调节的关系 纤溶抑制物活性，即“平衡”是如何调节的？ (源自上述实验的数据)；四)确定 由单核细胞形成的尿激酶原具有纤溶活性或 酶原(直接测定125I-纤溶酶原裂解以避免 尿蛋白原激活纤溶酶对尿蛋白的混杂作用。西式 印迹反应混合物以确定活性(PRO)UK为1或2 链)；以及v)确定单核细胞与尿激酶结合的效果 对其生物活性(作为纤溶酶原激活剂的效率，以及 对UK-I的易感性)。

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).