Molecular analysis of the adaptive immune response to tuberculosis

结核病适应性免疫反应的分子分析

• 批准号: 9204642
• 负责人: Spyros A Kalams
• 金额: $ 23.41万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-05 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204642
• 关键词: Affect; Antigens; Cell Line; Cell Separation; Cells; Complex; Defect; Epitope Mapping; Epitopes; Flow Cytometry; Immune; Immune response; Immune system; Immunology; In Vitro; Individual; Infection; Lung; Maps; Molecular; Molecular Analysis; Molecular Immunology; Mycobacterium tuberculosis; Peptides; Peripheral Blood Mononuclear Cell; Persons; Population; Production; Pulmonary Tuberculosis; Reporter; Residual state; Sorting - Cell Movement; Specificity; T cell response; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Tuberculosis; adaptive immunity; alpha-beta T-Cell Receptor; cohort; cytokine; exhaustion; immune activation; insight; latent infection; macrophage; pathogen; reconstitution; repository; research study; response

PROJECT SUMMARY The purpose of this R21 is to apply cutting edge molecular immunology techniques to determine the features of the adaptive immune response likely to protect against active tuberculosis infection. Although approximately 1/3 of the world’s population is infected with tuberculosis, the vast majority of infected individuals maintain control of latent TB infections throughout their lives. We have demonstrated in prior studies that individuals with prior treated extrapulmonary TB, and no sign of residual or new infection, have increased levels of immune activation compared to individuals with treated pulmonary TB, or treated latent infection. Furthermore, these individuals have lower levels of in vitro cytokine production in response to infected macrophages. This suggests an underlying immune defect in individuals prone to disseminated TB infection. We have established cutting edge molecular techniques that allow us to 1) single-cell sort pathogen-reactive T cells 2) directly sequence their T cell receptor (TCR) alpha and beta chains 3) determine the T cell receptor alpha and beta chains of each T cell and 4) Reconstitute the alpha beta TCR complex in a reporter cell line to allow fine mapping of epitope specificity. As a proof of concept, in this application we will apply these techniques to an existing repository of peripheral blood mononuclear cells from individuals with latent tuberculosis (treated and untreated) and individuals with prior treated pulmonary or extrapulmonary TB. We hypothesize that the degree of TB-specific TCR diversity may be related to the degree of control an individual has over latent infection.

项目摘要 该R21的目的是应用尖端分子免疫学技术来确定 适应性免疫反应可能预防活性结核病感染。虽然大约 世界人口中有1/3感染了结核病，绝大多数感染者维持 控制潜在结核病感染一生。我们在先前的研究中已经证明了 先前治疗的肺外结核病，没有残留或新感染的迹象，免疫水平升高 与接受治疗的肺结核或治疗的潜在感染相比，激活。此外，这些 响应感染的巨噬细胞，个体的体外细胞因子产生水平较低。这 表明在易于传播结核病感染的个体中的潜在免疫障碍。我们已经建立了 最先进的分子技术，使我们能够达到1）单细胞排序病原体反应性T细胞2）直接 将其T细胞受体（TCR）α和β链序列序列3）确定T细胞受体α和β 每个T细胞的链和4）重建记者细胞系列中的alpha beta TCR复合物以允许罚款 表位特异性的映射。作为概念的证明，在本应用程序中，我们将将这些技术应用于 来自潜在结核病患者的外周血单核细胞的现有存储库（治疗和 未经治疗的）和先前治疗的肺或肺外结核病患者。我们假设该学位 TB特异性的TCR多样性可能与个人对潜在感染的控制程度有关。