IMMUNODOMINANT ANTIGENS OF MYCOBACTERIUM TUBERCULOSIS

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，它在美国的发病率正在增加，因为 感染艾滋病毒的患者特别容易感染结核病。 根除结核病需要发展一种有效的 疫苗，这反过来又取决于对疫苗机制的理解 对结核病的保护性免疫。一系列的表现 结核病感染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 保护性免疫与迟发型超敏反应的相关性 以及对细胞内病原体的无效免疫力。这样做的目的是 建议是对T细胞亚群进行表征 结核保护性免疫与分枝杆菌的鉴定 能引起保护性免疫的抗原。这些抗原是潜在的 抗结核疫苗和皮肤诊断性测试的候选对象 试剂。我们的具体目标是：1)鉴定T细胞亚群 介导对分枝杆菌的抵抗性和保护性免疫反应 肺结核。主要的α-βT细胞抗原受体亚型 所有结核病患者的组织和血液 感染将通过用单抗免疫标记来识别 试剂结合聚合酶链式反应扩增α-βT细胞受体 基因。2)纯化和重组分枝杆菌的鉴定 通过α-βT细胞亚群诱导增殖的抗原 调节抵抗性和保护性免疫反应。抗原将会是 通过T细胞系筛选免疫反应性，并由以下定义的表位 针对合成肽测试T细胞克隆。免疫显性 抗原将通过有限稀释分析和 评估多种人类白细胞抗原类型间的抗原免疫反应性。3) 免疫优势抗原诱导人类免疫反应能力的研究 淋巴因子表型的功能反应 来自各种疾病患者的刺激T淋巴细胞 肺结核感染。将测量淋巴因子浓度 直接在细胞上清液中表达，并用聚合酶链式反应检测淋巴因子基因的表达 放大。4)免疫优势型结核分枝杆菌的评价 抗原作为诊断皮试试剂用于检测患者的皮肤试验 肺结核感染。用结核菌素中的这些抗原进行皮肤测试- 结核菌素阳性和阴性个体，结合测量 对淋巴细胞增殖的影响，将允许评估 体外免疫反应与体内诱导迟发型超敏反应的关系。

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.