Antiphospholipid Antibody Heterogeneity

抗磷脂抗体异质性

• 批准号: 6680671
• 负责人: NEAL STEWART ROTE
• 金额: $ 33.16万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6680671
• 关键词: SDS polyacrylamide gel electrophoresis; antiantibody; antibody specificity; antigen antibody reaction; antiphospholipid syndrome; apoptosis; autoantibody; binding proteins; blood chemistry; clinical research; enzyme linked immunosorbent assay; flow cytometry; fluorescence microscopy; human subject; immunoglobulin G; immunoglobulin M; ion exchange chromatography; medical complication; molecular cloning; phospholipids; polymerase chain reaction; pregnancy loss; protein purification; single cell analysis; trophoblast; western blottings

DESCRIPTION (provided by applicant): The Antiphospholipid Syndrome is characterized by naturally occurring antiphospholipid antibodies and increased risk of pregnancy loss or a variety of obstetric complications. Antiphospholipid antibodies are directed against phospholipid-dependent antigens, phospholipid-binding proteins, or a combination of these. There is little information concerning the relevance and relationship of particular antibody specificities to pathophysiology. Considerable data support the proposal that the placental trophoblast is a primary cellular target of antiphospholipid antibodies. The hypothesis is that antiphospholipid antibody-induced pregnancy loss occurs through the binding of a select subgroup of antibodies directly with trophoblast resulting in detrimental effects on placental development. Some of these effects involve thrombosis on the trophoblast surface, but most are non-thrombotic and result in incomplete trophoblast differentiation. A prediction, based on preliminary data, is that the most relevant antibodies will be against phosphatidylserine-dependent antigens. To test this hypothesis, the following Aims will be addressed. Aim 1 will investigate the heterogeneity of antiphospholipid antibodies within individual patients. This Aim is designed to determine within individual pregnancy loss patients the spectrum of reactivities against relevant phospholipids and phospholipid-binding proteins. Individual antiphospholipid antibodies will be purified based on antigenic specificity and characterized. Aim 2 will clone antiphospholipid antibodies from individual patients using single cell PCR techniques. The goal is to clone each relevant antibody present in a minimum of 3 individual patients. The cloned antibodies will be characterized. Aim 3 will investigate the relationship between antibody specificity and biological activity against trophoblast. Reagents prepared in Aims I and 2 will be tested for reactivity against trophoblast. These reactivities will include binding to normal human placental tissue, binding to trophoblast undergoing differentiation in vitro, inhibition of trophoblast invasion, inhibition of intertrophoblast fusion, and induction of apoptosis. This will be the first study to thoroughly investigate the spectrum of antiphospholipid antibodies in individual patients and correlate those data to effects on placental trophoblast.

描述（由申请人提供）：抗磷脂综合征的特征是自然产生的抗磷脂抗体和妊娠丢失或各种产科并发症的风险增加。抗磷脂抗体是针对磷脂依赖抗原，磷脂结合蛋白，或这些的组合。关于特定抗体特异性与病理生理的相关性和关系的信息很少。大量数据支持胎盘滋养细胞是抗磷脂抗体的主要细胞靶点的建议。该假说认为，抗磷脂抗体通过与滋养细胞直接结合的抗体亚群导致胎盘发育受损而导致妊娠丢失。其中一些作用涉及滋养层表面的血栓形成，但大多数作用是非血栓性的，并导致滋养层分化不完全。根据初步数据预测，最相关的抗体将是针对磷脂酰丝氨酸依赖抗原的抗体。为了验证这一假设，将解决以下目标。目的1将研究个体患者体内抗磷脂抗体的异质性。本目的目的是确定个体妊娠丢失患者对相关磷脂和磷脂结合蛋白的反应谱。个体抗磷脂抗体将根据抗原特异性进行纯化和鉴定。目的2将使用单细胞PCR技术从个体患者中克隆抗磷脂抗体。目标是克隆至少3个患者体内存在的每个相关抗体。克隆抗体将被表征。目的3将研究抗体特异性与抗滋养细胞生物活性之间的关系。在目标1和目标2中制备的试剂将测试对滋养细胞的反应性。这些反应性包括与正常人胎盘组织结合，与体外分化的滋养细胞结合，抑制滋养细胞侵袭，抑制滋养细胞间融合，诱导细胞凋亡。这将是第一个彻底调查个体患者抗磷脂抗体谱并将这些数据与胎盘滋养细胞的影响联系起来的研究。