CONTROL OF FIBRIN TURNOVER IN PLEURAL DISEASE

描述：(改编自申请人的摘要。)尽管很长时间 对胸膜纤维蛋白沉积与胸膜病变关系的认识 胸腔积液的组织情况，目前仅有有限的信息 关于胸膜纤维蛋白代谢紊乱的分子基础 受伤。人类渗出性胸腔积液被发现是 以促凝剂增加和纤溶抑制为特征 对这些胸腔积液细胞的活性和初步研究表明 间皮细胞在纤维蛋白沉积中起关键作用。 拟议的研究将解决以下假设：间皮细胞 促进纤维蛋白代谢异常，促进纤维蛋白生成 胸膜腔内的沉积。其作用机制及调控 间皮细胞和胸膜的促凝血和纤溶活性 体液将被确定。这些实验将涉及凝聚 纤溶酶原激活剂的分析、功能和免疫分析， 纤溶酶原激活物抑制物及活性物质的表征和检测 抗纤溶酶。 人胸膜间皮细胞可组装促凝剂 它们表面的复合体。调控的分子机制 组装外源性激活复合体(与组织因子相关的 因子VII/VIIa)和凝血酶原酶复合体(因子Xa与 将确定间皮细胞表面的Va)。约束性研究 将确定结合参数(Kd和位点数)、阳离子 外在激活的要求和功能活性 复合体和凝血酶原酶复合体。抗因子的单抗 VII和X将被用来验证结合的分子的数量和 将结合与功能活动联系起来。此外，结合部位 将使用一系列合成的多肽来探索特异性 与因子VII和因子X竞争。 由于间皮细胞功能纤溶活性低， 因为它们分泌高水平的纤溶酶原激活物抑制物 (PAI-1)，我们怀疑间皮细胞的PAI-1允许纤维蛋白。 证词。人培养物中PAI-1的调控机制 将对间皮细胞进行测定。这些实验将集中在 不同条件下纤溶酶原激活物-1基因的表达及其持续时间的研究 条件，如细胞与细胞因子的相互作用和 细胞外基质。

DESCRIPTION: (Adapted from the applicant's abstract.) Despite long standing recognition of the relation between pleural fibrin deposition and organization of pleural exudates, there is only limited information concerning the molecular basis of disordered fibrin turnover in pleural injury. Exudative pleural effusions in humans were found to be characterized by increased procoagulant and depressed fibrinolytic activities and pilot studies with cells from these pleural fluids suggest that the mesothelium plays a key role in fibrin deposition. Proposed studies will address the hypothesis that mesothelial cells contribute to abnormalities of fibrin turnover and promote fibrin deposition within the pleural space. The mechanisms and regulation of procoagulant and fibrinolytic activity in mesothelial cells and pleural fluids will be determined. These experiments will involve coagulation assays, functional and immunologic assays of plasminogen activators, characterization and assay of plasminogen activator inhibitors and antiplasmins. It was found that human pleural mesothelial cells assemble procoagulant complexes on their surface. The molecular mechanisms that regulate assembly of the extrinsic activation complex (tissue factor associated with factor VII/VIIa) and the prothrombinase complex (factor Xa associated with Va) at the mesothelial cell surface will be determined. Binding studies will determine binding parameters (Kd and number of sites), cation requirements and functional activity of both the extrinsic activation complex and the prothrombinase complex. Monoclonal antibodies to factors VII and X will be used to verify the number of molecules bound and to relate binding with functional activity. In addition, binding site specificity will be probed using a series of synthetic peptides that compete with factors VII and X. Because mesothelial cells have low functional fibrinolytic activity, and because they secrete high levels of plasminogen activator inhibitor (PAI-1), we suspect that PAI-1 from the mesothelium permits fibrin deposition. The mechanisms that regulate PAI-1 in cultures of human mesothelial cells will be determined. These experiments will focus on the expression of mRNA for PAI-1 and the duration of the message under various conditions, such as cellular interaction with cytokines and the extracellular matrix.