Development &validationof a panel of antibodies for the diagnosis of sarcoidosis

描述（由申请方提供）：本提案将检验免疫在结节病发病机制中起重要作用的假设，如免疫功能异常和抗体存在所示，包括在该疾病中发现的类风湿因子和抗核抗体。此外，结节病与其他全身性炎症性疾病共存，患者常表现为皮肤无反应性和高丙种球蛋白血症。我们建议开发一组具有高灵敏度和特异性的抗体/自身抗体，以识别结节病患者，并有助于将这些患者与对照组和其他肺部疾病患者区分开来。为此，我们将构建一个T7噬菌体的潜在结节病抗原的cDNA文库，使用从结节病患者的受影响的淋巴结组织中分离的mRNA。该cDNA文库将用含有高滴度IgG抗体的结节病患者血清进行免疫筛选，克隆的菌落将用于构建抗原微阵列，该抗原微阵列将与肺结节病病例和对照的血清杂交。对照血清将从血清阳性类风湿性关节炎患者和有结核病史的患者中获得。通过对结节病患者血清识别为抗原的噬菌体插入片段进行序列分析，可以鉴定与结节病发病机制相关的抗原。将使用从病例和对照中独立获得的血清样本对自身抗体分类器进行验证。在未来，这种方法可能会确定用于治疗结节病的分子靶点。

DESCRIPTION (provided by applicant): This proposal will test the hypothesis that immunity has a prominent role in the pathogenesis of sarcoidosis, as suggested by abnormalities of the immune function and by the presence of antibodies, including rheumatoid factors and antinuclear antibodies found in this disorder. In addition, sarcoidosis coexists with other systemic inflammatory diseases, and patients frequently show cutaneous anergy and hypergammaglobulinemia. We propose to develop a panel of antibodies/autoantibodies with high sensitivity and specificity to identify patients with sarcoidosis and help to distinguish these patients from controls and individuals with other pulmonary diseases. For this purpose we will construct a T7 phage cDNA library of potential sarcoidosis antigens using mRNA isolated from affected lymph node tissue of patients with sarcoidosis. This cDNA library will be immunoscreened with sera from patients with sarcoidosis containing high titer IgG antibodies and the cloned colonies will be used to construct an antigen microarray that will be hybridized with sera from cases with lung sarcoidosis and controls. Control sera will be obtained from patients with sero-positive rheumatoid arthritis and from patients with a history of tuberculosis. Sequence analyses of informative phage inserts recognized as antigens by sarcoidosis patient sera may identify antigens associated with the etiopathogenesis of sarcoidosis. The autoantibody classifier will be validated with independently obtained collections of sera from cases and controls. In the future, this approach may identify molecular targets useful for the treatment of sarcoidosis.