IMMUNODOMINANT ANTIGENS OF MYCOBACTERIUM TUBERCULOSIS

结核分枝杆菌的免疫显性抗原

• 批准号: 2063788
• 负责人: Peter F. Barnes
• 金额: $ 25.27万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-02-01 至 1996-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2063788
• 关键词: Mycobacterium tuberculosis; T cell receptor; bacterial antigens; cellular immunity; cytokine; genetic library; helper T lymphocyte; human tissue; lymphocyte proliferation; monoclonal antibody; polymerase chain reaction; tissue /cell culture; tuberculosis; tuberculosis vaccines

Tuberculosis causes more deaths world-wide than any other infectious pathogen, and its incidence is increasing in the United States because HIV-infected patients are extraordinarily susceptible to tuberculosis. Eradication of tuberculosis requires development of an effective vaccine, which in turn hinges on understanding the mechanism of protective immunity against tuberculosis. The spectrum of manifestations of tuberculosis infection reflects the cell-mediated immune response, and provides an excellent model to characterize the immunologic correlates of protective immunity, delayed-type hypersensitivity (DTH) and ineffective immunity to intracellular pathogens. The goals of this proposal are to characterize T cell subpopulations that mediate protective immunity against tuberculosis and to identify mycobacterial antigens that elicit protective immunity. These antigens are potential candidates for an antituberculosis vaccine and for diagnostic skin test reagents. Our specific aims are: 1) Identification of T cell subsets that mediate resistant and protective immune responses to Mycobacterium tuberculosis. Dominant alpha beta T cell antigen receptor subtypes in tissue and blood of patients throughout the spectrum of tuberculosis infection will be identified by immunolabeling with monoclonal reagents, combined with PCR amplification of alpha beta T cell receptor genes. 2) Identification of purified and recombinant mycobacterial antigens that elicit proliferation by alpha beta T cell subpopulations that mediate resistant and protective immune responses. Antigens will be screened for immunoreactivity by T cell lines, and epitopes defined by testing T cell clones against synthetic peptides. Immunodominant antigens will be identified by limiting dilution analysis and by evaluating immunoreactivity of antigen across multiple HLA types. 3) Investigation of the capacity of immunodominant antigens to induce a functional response by characterization of the lymphokine profile of stimulated T-lymphocytes from patients across the spectrum of tuberculosis infection. Lymphokine concentrations will be measured directly in cell supernatants, and lymphokine mRNA detected by PCR amplification. 4) Evaluation of immunodominant M. tuberculosis-specific antigens as diagnostic skin test reagents to detect patients with tuberculosis infection. Skin testing with these antigens in tuberculin- positive and tuberculin-negative individuals, combined with measurement of lymphocyte proliferation, will allow assessment of the relationship between immunoreactivity in vitro and induction of DTH in vivo.

结核病在世界范围内造成的死亡人数比任何其他传染病都多。 病原体，其发病率在美国正在增加，因为 艾滋病毒感染者特别容易感染结核病。 根除结核病需要制定有效的 疫苗，这反过来又取决于了解的机制， 对结核病的保护性免疫。表现的范围 反映了细胞介导的免疫反应， 并提供了一个很好的模型来表征免疫学 保护性免疫、迟发型超敏反应（DTH） 以及对细胞内病原体的无效免疫。这个的目标 建议是表征T细胞亚群，介导 对结核病的保护性免疫和鉴定分枝杆菌 引发保护性免疫的抗原。这些抗原是潜在的 抗结核疫苗和诊断性皮肤试验的候选者 试剂我们的具体目标是：1）识别T细胞亚群 介导对分枝杆菌的抵抗性和保护性免疫反应 结核T细胞抗原受体亚型的研究进展 所有结核病患者的组织和血液 感染将通过用单克隆抗体进行免疫标记来鉴定。 试剂，结合PCR扩增α β T细胞受体 基因. 2)纯化和重组分枝杆菌的鉴定 引发α β T细胞亚群增殖的抗原 介导抵抗性和保护性免疫反应。抗原将是 通过T细胞系筛选免疫反应性， 测试针对合成肽的T细胞克隆。免疫显性 抗原将通过有限稀释分析和 评估多种HLA类型的抗原的免疫反应性。第三条 研究免疫显性抗原诱导a 功能反应通过表征的淋巴因子概况 刺激的T淋巴细胞从患者跨越频谱 肺结核感染。将测量细胞因子浓度 直接在细胞上清液中，并通过PCR检测淋巴因子mRNA 放大4)免疫优势M.结核病特异性 抗原作为诊断性皮肤试验试剂， 肺结核感染。在结核菌素中用这些抗原做皮肤试验- 阳性和结核菌素阴性个体，结合测量 淋巴细胞增殖，将允许评估的关系 在体外的免疫反应性和在体内的DTH诱导之间。