ANTIPHOSPHOLIPID ANTIBODIES AND DISORDERS OF PREGNANCY

抗酸性磷脂（aPL）的自身抗体与 血栓形成和反复流产。 虽然杀伤人员地雷被认为 通过不适当的凝血激活直接导致这些疾病 这一假设没有得到任何公开数据的明确支持或反驳。 同样，对有关的问题也有相当多的辩论。 杀伤人员地雷的特殊性及其作用机制， 个体患者，具有明显相同的aPL特异性， 在症状上有很大的差异。 我们的中心假设是， 直接导致流产 我们的初步数据 进一步引导我们提出三个可检验的具体假设。 第一 aPL在特异性上是异源的， 确定特定患者临床症状的严重程度。 的 第二个假设是三种细胞类型可能是 aPL;血管内皮细胞、血小板和滋养层。 这些细胞中的哪一个主要受到影响取决于集体 特定患者的APL的特异性。 第三个假设是 杀伤人员地雷将直接导致实验动物生殖失败， 动物 为了验证这些假设，我们提出了五个具体目标。 设计了 首先，使用单克隆抗体和来自 我们将评估aPL的异质性， ELISA研究，与已知磷脂的吸附，与 人造膜，磷脂亲和柱上的洗脱曲线， 和代表的独特类型的比较。 第二，我们将调查 aPL对血小板活化的影响。 使用流式细胞术和 抗血小板特异性抗原单克隆抗体及血小板 活化抗原，以及血栓素的体外测量 生产，我们将测量aPLs对静息和激活的反应性。 血小板，确定aPL是否直接激活血小板或增强 血小板对其他激活剂的反应，并评估 原位血小板活化。 第三，我们将评估反应性， aPL对培养的牛和人内皮细胞的影响，包括 它们与静息和活化的内皮细胞的结合，它们对 内皮细胞代谢和内皮细胞的表征 与aPL反应的抗原。 第四，使用胎盘制剂， 分离的细胞滋养层细胞和绒毛膜癌细胞系BeWo， 研究aPL的结合和作用，包括对体外 细胞滋养层细胞间融合和总体代谢活性。 第五，使用怀孕的小鼠和豚鼠，我们将测试直接 杀伤人员地雷对流产的生物学影响。 这项研究的成果 将在体外和体内分析aPL的特异性和功能， 与流产的关系

Autoantibodies against acidic phospholipids (aPLs) are associated with thrombosis and recurrent pregnancy loss. Although aPLs are believed to directly cause these disorders through inappropriate activation of clotting this assumption is not clearly supported or refuted by any published data. Likewise, there is considerable debate concerning the relevant specificities of aPLs and their mechanisms of action, as well as why individual patients, with apparently identical aPL specificities, present with a large difference in symptoms. Our central hypothesis is that aPLs are directly responsible for pregnancy loss. Our preliminary data would further lead us to propose three testable specific hypotheses. The first is that aPLs are heterologous in specificity, the nature of which determines the severity of the particular patient's clinical symptoms. The second hypothesis is that three cell types may be principal targets of aPLs; the vascular endothelial cell, the platelet, and the trophoblast. Which of these cells is primarily affected depends upon the collective specificities of that particular patient's aPLs. The third hypothesis is that aPLs will directly induce reproductive failure in experimental animals. In order to test these hypotheses five specific aims have been designed. First, using monoclonal antibodies and purified antibodies from patients we will evaluate the heterogeneity of aPLs through blocking studies in ELISAs, adsorption with known phospholipids, reactivity with artificial membranes, elution profiles on phospholipid affinity columns, and comparison of represented idiotypes. Second, we will investigate the effects of aPLs on platelet activation. Using flow cytometry and monoclonal antibodies against platelet specific antigens and platelet activation antigens, as well as in vitro measurement of thromboxane production, we will measure the reactivity of aPLs on resting and activated platelets, determine whether aPLs directly activate platelets or enhance the response of platelets to other activators, and evaluate the level of platelet activation in situ. Third, we will evaluate the reactivity and effects of aPLs on cultured bovine and human endothelial cells, including their binding to resting and activated endothelial cells, their effect on endothelial cell metabolism, and the characterization of endothelial cell antigens reactive with aPLs. Fourth, using placental preparations, isolated cytotrophoblast, and a choriocarcinoma line, BeWo, we will investigate the binding and effect of aPLs, including effects on in vitro cytotrophoblast intercellular fusion and overall metabolic activity. Fifth, using pregnant mice and guinea pigs we will test the direct biological effects of aPLs on pregnancy loss. The product of this study will be an in vitro and in vivo analysis of aPL specificity and function in relationship to pregnancy loss.