AUTOANTIBODIES IN SCLERODERMA AND RAYNAUD'S PHENOMENON

CREST患者血清中存在的抗着丝粒抗体（ACA）或 雷诺现象伴或不伴结缔组织疾病或 雷诺氏病已被发现与3个着丝粒蛋白反应 （CENP）：CENP-A、CENP-B和CENP-C。 CENP-B似乎是主要的 自身抗原已被克隆并在E.杆菌 抗Scl-70抗体 已显示与拓扑异构酶I反应，拓扑异构酶I也已被克隆。 使用 免疫印迹技术，抗拓扑异构酶I可在血清中显示 免疫扩散检测抗SCL-70阴性。 初步数据提示 抗拓扑异构酶I的存在可以预测 硬皮病和CENP-B抗体可预测CREST发生 综合征 3种着丝粒蛋白和拓扑异构酶抗体的存在 我将通过染色体免疫印迹进行研究。 水平 抗CENP-B和拓扑异构酶I将通过ELISA测定使用 融合蛋白CENP-B和拓扑异构酶I。 我们假设，有一个共同的遗传背景的患者， 这些自身抗体可以通过寻找 交叉反应性独特型 亲和纯化的CENP-B自身抗体 以及针对CENP-B和拓扑异构酶I的自身抗体 人-人杂交瘤产生的多克隆抗体将用于产生 兔抗独特型和小鼠单克隆抗独特型。 我们将 寻找患者血清中的交叉反应独特型， 亲戚 将使用以下方法研究含有交叉反应独特型的血清： 电聚焦和标记的抗原，以确定是否交叉 反应性独特型与抗原反应。 一项对雷诺现象患者的前瞻性研究， 将进行结缔组织疾病的发作，以确定 抗CENP-B的存在和量的临床意义， 拓扑异构酶I 此外，存在的预后意义 抗CENP-A和抗CENP-C的抗体将被测定。

Anticentromere antibodies (ACA) present in sera from patients with CREST or Raynaud's phenomenon with or without a connective tissue disease or in Raynaud's disease have been found to react with 3 centromeric proteins (CENPs): CENP-A, CENP-B and CENP-C. CENP-B which appears to be the major autoantigen has been cloned and is expressed in E. coli. Anti-Scl-70 has been shown to react with topoisomerase I which has also been cloned. Using immunoblotting technique, anti-toposomerase I can be demonstrated in sera negative for anti-SCL-70 on immunodiffusion. Preliminary data suggests that the presence of anti-topoisomerase I may predict the development of scleroderma and that anti-CENP-B may predict the development of CREST syndrome. The presence of antibodies to the 3 centromeric proteins and topoisomerase I will be studied by immunoblotting with chromosomes. The level of anti-CENP-B and topoisomerase I will be studied by ELISA assay using the fusion protein CENP-B and topoisomerase I. We postulate that there is a common genetic background in patients with these autoantibodies which can be demonstrated by looking for the presence of cross reactive idiotypes. Affinity purified auto-antibodies to CENP-B and to topoisomerase I and autoantibodies to CENP-B and topoisomerase I generated by human-human hybridomas will be used to produce polyclonal anti-idiotypes in rabbits and monoclonal anti-idiotypes in mice. We will look for cross reactive idiotypes in sera from patients and their relatives. Sera containing cross reactive idiotype will be studied using electrofocusing and labelled antigen to determine whether or not the cross reactive idiotype reacts with antigen. A prospective study of patients with Raynaud's phenomenon unassociated at the onset with a connective tissue disease will be carried out to determine the clinical significance of the presence and amount of anti-CENP-B and topoisomerase I. In addition, the prognostic significance of the presence of anti-CENP-A and anti-CENP-C will be determined.