Antiphospholipid Antibody Heterogeneity

抗磷脂抗体异质性

• 批准号: 6881648
• 负责人: NEAL STEWART ROTE
• 金额: $ 34.09万
• 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/6881648
• 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天然发生的抗磷脂抗体和妊娠丧失或多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。大量数据支持以下提议：胎盘滋养细胞是抗磷脂抗体的主要细胞靶标。假设是，抗磷脂抗体诱导的妊娠丧失是通过直接与滋养细胞的精选亚组结合的结合，从而对胎盘发育产生不利影响。这些作用中的一些涉及滋养细胞表面的血栓形成，但大多数是非栓性的，导致滋养细胞分化不完全。基于初步数据的预测是，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。 AIM 1将研究个别患者内抗磷脂抗体的异质性。该目的旨在确定个体妊娠损失患者的反应率对相关的磷脂和磷脂结合蛋白的频谱。单个抗磷脂抗体将根据抗原特异性纯化并表征。 AIM 2将使用单细胞PCR技术从单个患者中克隆抗磷脂抗体。目的是克隆至少3名单独患者的每种相关抗体。克隆的抗体将被表征。 AIM 3将研究抗体特异性与针对滋养细胞的生物活性之间的关系。在目标I和2中制备的试剂将进行针对滋养细胞的反应性。这些反应率将包括与正常的人胎盘组织结合，与受分化的滋养细胞结合，抑制滋养细胞侵袭，抑制人际化质细胞融合以及诱导细胞凋亡。这将是第一个彻底研究个别患者抗磷脂抗体的频谱并将这些数据相关联以影响胎盘滋养细胞的研究。