Antiphospholipid Antibody Heterogeneity

抗磷脂抗体异质性

• 批准号: 6765913
• 负责人: NEAL STEWART ROTE
• 金额: $ 33.39万
• 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/6765913
• 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）：抗磷脂综合症的特征是天然存在的抗磷脂抗体以及流产或各种产科并发症的风险增加。抗磷脂抗体针对磷脂依赖性抗原、磷脂结合蛋白或它们的组合。 关于特定抗体特异性与病理生理学的相关性和关系的信息很少。大量数据支持胎盘滋养层是抗磷脂抗体的主要细胞靶标的观点。该假设认为，抗磷脂抗体诱导的妊娠丢失是通过选定的抗体亚组直接与滋养层结合而发生的，从而对胎盘发育产生有害影响。其中一些影响涉及滋养层表面的血栓形成，但大多数是非血栓形成的并导致滋养层分化不完全。根据初步数据预测，最相关的抗体将针对磷脂酰丝氨酸依赖性抗原。为了检验这一假设，将解决以下目标。目标 1 将研究个体患者内抗磷脂抗体的异质性。该目的旨在确定个体妊娠丢失患者针对相关磷脂和磷脂结合蛋白的反应谱。各个抗磷脂抗体将根据抗原特异性进行纯化并进行表征。目标 2 将使用单细胞 PCR 技术克隆个体患者的抗磷脂抗体。目标是克隆至少 3 名患者体内存在的每种相关抗体。克隆的抗体将被表征。目标 3 将研究抗体特异性和针对滋养层的生物活性之间的关系。将测试目标 I 和 2 中制备的试剂针对滋养层的反应性。这些反应性包括与正常人胎盘组织结合、与体外分化的滋养层结合、抑制滋养层侵袭、抑制滋养层间融合以及诱导细胞凋亡。这将是第一项彻底研究个体患者抗磷脂抗体谱并将这些数据与对胎盘滋养层的影响相关联的研究。