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

发展

• 批准号: 8243195
• 负责人: Lobelia Samavati
• 金额: $ 18.22万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8243195
• 关键词: Affect; African American; Antibodies; Antigens; Antinuclear Antibodies; Autoantibodies; Bacterial Antigens; Bacteriophage T7; Bacteriophages; Clinical; Collection; Crohn' s disease; Cutaneous; Databases; Development; Diagnosis; Diagnostic; Disease; Drug Delivery Systems; Etiology; Eye; Future; Granuloma; Granulomatous; Health; Humoral Immunities; Hypergammaglobulinemia; Immune System Diseases; Immune response; Immunity; Immunoglobulin G; Immunotherapy; Incidence; Individual; Inflammation; Inflammatory; Lung; Lung diseases; Lupus Erythematosus; Lymph Node Tissue; Mediastinal lymph node group; Messenger RNA; Molecular Target; Neuraxis; Organ; Pathogenesis; Pathway interactions; Patients; Pulmonary Sarcoidosis; Recording of previous events; Rheumatoid Arthritis; Rheumatoid Factor; Role; Sarcoidosis; Sensitivity and Specificity; Sequence Analysis; Sequence Homology; Serum; Skin; Societies; Testing; Tissues; Tuberculosis; Viral Antigens; Woman; anergy; base; cDNA Library; case control; human disease; immune function; instrument; lymph nodes; men; tool

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.

描述（由申请人提供）：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，正如免疫功能异常和抗体（包括在这种疾病中发现的类风湿因子和抗核抗体）的存在所表明的那样。此外，结节病与其他全身炎症性疾病并存，患者常表现出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体区分开来。为此，我们将使用从结节病患者受影响的淋巴结组织中分离的 mRNA 构建潜在结节病抗原的 T7 噬菌体 cDNA 文库。该 cDNA 文库将使用来自结节病患者的含有高效价 IgG 抗体的血清进行免疫筛选，克隆的集落将用于构建抗原微阵列，该抗原微阵列将与来自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。对被结节病患者血清识别为抗原的信息丰富的噬菌体插入序列进行序列分析，可以鉴定与结节病发病机制相关的抗原。自身抗体分类器将使用从病例和对照中独立获得的血清集合进行验证。将来，这种方法可能会识别出可用于治疗结节病的分子靶点。 公共卫生相关性：异常的免疫反应是多种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，与克罗恩病、类风湿性关节炎和红斑狼疮等许多炎症或肉芽肿性疾病有相似之处。结节病在世界各地都有发生，男性的平均发病率为 16.5/100,000，女性的平均发病率为 19/100,000，但非洲裔美国人的发病率高达 75/100,000。由于其长期性以及影响年轻人的趋势，它对我们社会的健康和经济产生了重大影响。自从一个多世纪前描述这种情况以来，研究未能确定导致结节病肉芽肿性炎症的特定抗原。拟议的研究将鉴定一组识别结节病肉芽肿抗原的抗体。这将可作为诊断肺结节病的工具，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一种可用于临床诊断肺结节病的诊断仪器。信息丰富的噬菌体的表征可能揭示与肺结节病发展相关的细菌或病毒抗原的存在。该提案的长期目标是鉴定器官特异性抗原，这些抗原可以作为药物的靶标或作为免疫治疗的基础。