CONTROL MECHANISMS IN HEMOSTASIS

这项建议的目的包括3个主要目标： 1.鉴定FVIII介导的血小板黏附的性质 血管内皮下层。我们将比较FVIII和Fviii的约束力 去内皮FVIII(ASVIII)对兔和人血管的作用， 培养内皮细胞，内皮细胞细胞外基质，胶原 和微纤维，以及FVIII和ASVIII对血小板的调节作用 在这些系统中的粘附性。我们还将ASVIII约束与 未刺激的血小板与刺激的FVIII结合的血小板 正常人群、阿司匹林正常人群和储藏池疾病 综合征和血栓减少患者。我们将研究其抑制作用 我们的抗血小板GPIb和ASVIII单抗与FVIII和ASVIII结合 GPIIb-IIIa复合体。 2.确定狼疮抗凝剂的作用机制(S)，以及 阴离子反应循环抗体的止血意义 磷脂。我们将继续对阴离子磷脂的研究 狼疮抗凝剂的特异性及其与抗DNA抗体的比较 狼疮患者的抗体。我们将研究对前列腺素I2的抑制作用 狼疮抗凝剂对培养内皮细胞合成的影响 经等电聚焦或亲和层析纯化 PS，PA-琼脂糖凝胶。我们将研究其机制。 低凝血酶原血症，27%的狼疮患者服用抗凝剂 狼疮患者与非狼疮患者的125I-凝血酶原转化率研究 抗凝剂。 3.探讨血小板与血管内皮细胞的关系 膜蛋白。我们将描述人类内皮细胞的特征 膜组分与单抗和多克隆抗体反应 血小板GpIIIa，确定该成分是否具有P1A1抗原性 特异性(如血小板GpIIIa)，并研究这些结合是否 抗体改变内皮细胞的功能，特别是PGI2的产生。 我们将研究血栓患者血清中的免疫球蛋白组份。 血小板减少性紫癜，可能有针对谁的抗体 内皮细胞和/或血小板，用于与内皮细胞的反应性 细胞，特别是针对GpIIIa成分。我们将进行搜索 对于内皮细胞和内皮细胞之间共有的其他膜成分 血小板。

The aims of this proposal cover 3 major objectives: 1. To characterize the nature of FVIII-mediated platelet adhesion to vascular subendothelium. We shall compare the binding of FVIII and desialylated FVIII (ASVIII) to deendothelialized rabbit and human vessels, cultured endothelial cells, endothelial cell extracellular matrix, collagen and microfibrils, and the effect of FVIII and ASVIII in modulating platelet adhesion in these systems. We shall also compare ASVIII binding to unstimulated platelets with FVIII binding to stimulated platelets from normals, aspirinized normals, and storage pool disease, Bernard-Soulier syndrome and thrombasthenic patients. We shall study the inhibition of FVIII and ASVIII binding by our monoclonal antibodies to platelet GpIb and the GpIIb-IIIa complex. 2. To determine the mechanism(s) of action of lupus anticoagulants, and the hemostatic significance of circulating antibodies reactive with anionic phospholipids. We shall continue studies of the anionic phospholipid specificity of lupus anticoagulants and compare them with anti-DNA antibodies from patients with lupus. We shall study inhibition of PGI2 synthesis in cultured endothelial cells produced by lupus anticoagulants purified by isoelectric focusing or affinity chromatography on PS,PA-Sepharose. We shall investigate the mechanism of hypoprothrombinemia, seen in 27% of patients with lupus anticoagulants by doing 125I-prothrombin turnover studies in lupus patients with and without anticoagulants. 3. To investigate the relationship between platelet and endothelial cell membrane proteins. We shall characterize the human endothelial cell membrane component reactive with monoclonal and polyclonal antibodies to platelet GpIIIa, determine whether this component has P1A1 antigenic specificity (like platelet GpIIIa), and study whether the binding of these antibodies alters endothelial cell function, particularly PGI2 production. We shall study IgG fractions from sera of patients with thrombotic thrombocytopenia purpura, who may have antibodies directed toward endothelial cells and/or platelets, for reactivity against endothelial cells, and against the GpIIIa component, in particular. We shall search for other membrane components in common between endothelial cells and platelets.