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。 与公共卫生相关：异常的免疫反应是导致大量人类疾病的主要原因。结节病是一种病因不明的炎症性疾病，与许多炎症性或肉芽肿性疾病如克罗恩病、类风湿性关节炎和红斑狼疮有相似之处。结节病在世界各地都有发生，男性的平均发病率为16.5/10万，女性为19/10万，但非洲裔美国人的发病率高达75/10万。由于它的长期性和影响年轻人的倾向，它对我们社会的健康和经济产生了很大的影响。自从一个多世纪前对这种情况的描述以来，研究一直未能确定导致结节病肉芽肿性炎症的特定抗原。拟议的研究将确定一组识别结节病肉芽肿抗原的抗体。这将是一种诊断肺结节病的有用工具，并可能确定治疗这种疾病的分子靶点。该项目的主要成果将是在临床环境中用于诊断肺结节病的诊断仪器。信息噬菌体的特性可能揭示与肺结节病的发展相关的细菌或病毒抗原的存在。这项提议的一个长期目标是确定器官特异性抗原，这些抗原可能是药物的靶标，也可能是免疫治疗的基础。

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. PUBLIC HEALTH RELEVANCE: Aberrant immune responses are a major cause of a vast array of human diseases. Sarcoidosis is an inflammatory disease of unknown etiology sharing similarities with many inflammatory or granulomatous diseases such as Crohn's disease, rheumatoid arthritis, and lupus erythematosus. Sarcoidosis occurs throughout the world with an average incidence of 16.5/100,000 in men and 19/100,000 in women but up to 75/100,000 in African-Americans. Due to its chronicity and to its tendency to affect young individuals, it has high impact on the health and economy in our society. Since the description of this condition more than a century ago, studies have failed to identify specific antigens leading to granulomatous inflammation in sarcoidosis. The proposed studies will identify a panel of antibodies recognizing sarcoidosis granuloma antigens. This will be useful as a tool to diagnose pulmonary sarcoidosis and may identify molecular targets for the treatment of this disease. The main delivery from this project will be a diagnostic instrument useful in a clinical setting for the diagnosis of pulmonary sarcoidosis. Characterization of informative phage may reveal the presence of antigens from bacterial or viral antigens related to the development of pulmonary sarcoidosis. A long term objective of this proposal is the identification of organ-specific antigens which could be targeted by drugs or be the basis for immunotherapy.