TREATMENT STRATEGIES FOR THE ANTI PHOSPHOLIPID SYNDROME

抗磷脂综合征的治疗策略

• 批准号: 6498969
• 负责人: JOHN R DEDMAN
• 金额: $ 25.4万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6498969
• 关键词: autoimmune disorder; cardiolipins; combinatorial chemistry; epitope mapping; genetically modified animals; immunotherapy; laboratory mouse; monoclonal antibody; nonhuman therapy evaluation; peptide library; phospholipids; syndrome; thrombosis

Antiphospholipid Syndrome (APS), also known as Hughes Syndrome, is a multiorgan vascular disease associated with myocardial infarction, stroke thrombosis and recurrent fetal loss. Forty- five percent of the people under the age of fifty who suffer strokes also demonstrate elevated levels of phospholipid antibody. Our hypothesis is that the presence of antiphospholipid antibodies in the circulation leads to vascular disorders, and that the escalating tissue damage can be abated with agents that bind the destructive antibodies. In order to test this hypothesis, it is necessary to create a genetically defined APS mouse that secretes antiphospholipid antibodies into the blood. We have produced and characterized a mouse monoclonal antibody which specifically recognizes phosphatidylserine. Preliminary data demonstrates that our monoclonal antibody binding specificity is similar to antibodies present in APS patients; this monoclonal antibody is therefore appropriate to develop a treatment model. The heavy and light chain of our monoclonal antiphospholipid antibody will be cloned by RT-PCR. These cDNAs will include the endogenous secretory peptide signal sequence. Bigenic mice will be used to direct ligand induced expression in the liver. Each monoclonal immunoglobulin chain will be differentially tagged with HA or FLAG epitopes. The transgenic animals will be characterized and compared with normal animals for the hallmark symptoms of APS, including prolonged in vitro coagulation times, recurrent fetal loss and vascular disease. An additional goal of this proposal is to develop treatment strategies for APS. Our approach is to neutralize the disease-causing antibodies. Peptides which bind and block the monoclonal antiphospholipid antibody in vitro can be selected from a combinatorial phage-display library of random peptides. Isolated peptides will be assessed for the ability to reduce the symptoms of APS displayed by this transgenic mouse. This study will produce a genetically-defined mouse model of APS which will prove useful for developing treatment strategies to ameliorate the symptoms associated with APS.

抗磷脂综合征（APS），也称为休斯综合征，是一种多器官血管疾病，与心肌梗死，中风血栓形成和复发性胎儿丢失有关。 在50岁以下中风的人中，45%的人也表现出磷脂抗体水平升高。 我们的假设是，抗磷脂抗体在循环中的存在会导致血管疾病，而不断升级的组织损伤可以用结合破坏性抗体的药物来减轻。 为了检验这一假设，有必要建立一个基因定义的APS小鼠分泌抗磷脂抗体进入血液。 我们已经产生并表征了特异性识别磷脂酰丝氨酸的小鼠单克隆抗体。初步数据表明，我们的单克隆抗体结合特异性与APS患者中存在的抗体相似;因此，该单克隆抗体适用于开发治疗模型。 我们的单克隆抗磷脂抗体的重链和轻链将通过RT-PCR克隆。这些cDNA将包括内源性分泌肽信号序列。 双基因小鼠将用于指导肝脏中的配体诱导表达。 每个单克隆免疫球蛋白链将用HA或FLAG表位进行差异标记。 将对转基因动物进行表征，并与正常动物比较APS的标志性症状，包括体外凝血时间延长、复发性流产和血管疾病。 该提案的另一个目标是制定APS的治疗策略。 我们的方法是中和致病抗体。 在体外结合和阻断单克隆抗磷脂抗体的肽可以选自随机肽的组合噬菌体展示文库。将评估分离的肽减轻该转基因小鼠表现出的APS症状的能力。 这项研究将产生一个遗传定义的APS小鼠模型，这将证明有助于开发治疗策略，以改善与APS相关的症状。