CONTROL OF FIBRIN TURNOVER IN PLEURAL DISEASE

描述：（改编自申请人的摘要。）尽管长 对胸膜纤维蛋白沉积与 胸腔渗出物的组织，信息有限 关于胸膜纤维蛋白转换紊乱的分子基础 损伤 人类的渗出性胸腔积液被发现是 以促凝血剂增加和纤溶抑制为特征 活动和对这些胸腔积液细胞的初步研究表明， 间皮细胞在纤维蛋白沉积中起着关键作用。 拟议的研究将解决这一假设，即间皮细胞 导致纤维蛋白周转异常， 胸膜腔内沉积物。 的机制和调控 间皮细胞和胸膜细胞中的促凝血和纤溶活性 液体将被确定。 这些实验将涉及凝血 纤溶酶原激活剂的测定、功能和免疫测定， 纤溶酶原激活物抑制剂的表征和测定， 抗纤溶酶。 发现人胸膜间皮细胞组装促凝血剂 复合物在其表面。 调节细胞凋亡的分子机制 外源性激活复合物（与之相关的组织因子 因子VII/VIIa）和凝血酶原酶复合物（因子Xa与 Va）在间皮细胞表面的浓度。 结合研究 将确定结合参数（Kd和位点数）、阳离子 需要和功能活动的外源性激活 复合物和凝血酶原酶复合物。 抗因子单克隆抗体 VII和X将用于验证结合的分子数量， 将结合与功能活性联系起来。 此外，结合位点 将使用一系列合成肽探测特异性， 与因子VII和因子X竞争。 由于间皮细胞具有低功能性纤溶活性， 因为它们分泌高水平的纤溶酶原激活物抑制剂 （派-1），我们怀疑来自间皮的派-1允许纤维蛋白 证词 派-1在人培养细胞中的调节机制 将测定间皮细胞。 这些实验将集中在 派-1的mRNA表达和在各种条件下的信息持续时间 条件，如细胞与细胞因子的相互作用，以及 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.