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

描述(申请人提供)：这项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的存在，包括在这种疾病中发现的类风湿因子和抗核抗体。此外，结节病与其他全身炎症性疾病并存，患者经常表现为皮肤无能和高丙种球蛋白血症。我们建议开发一组具有高敏感性和特异性的抗体/自身抗体来识别结节病患者，并帮助将这些患者与对照组和患有其他肺部疾病的个人区分开来。为此，我们将使用从结节病患者受影响的淋巴组织中提取的mRNA构建潜在的结节病抗原的T7噬菌体cDNA文库。该文库将与含有高效价免疫球蛋白抗体的结节病患者血清进行免疫筛选，克隆的克隆将用于构建与肺结节病患者和对照组血清杂交的抗原芯片。对照血清将从血清阳性类风湿性关节炎患者和有肺结核病史的患者那里获得。对被结节病患者血清识别为抗原的信息噬菌体插入片段进行序列分析，可以确定与结节病病因相关的抗原。该自身抗体分类器将通过从病例和对照中独立获得的血清收集来验证。在未来，这种方法可能会确定对治疗结节病有用的分子靶点。

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.