Molecular analysis of the adaptive immune response to tuberculosis

项目总结 此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会预防活动性结核病感染。尽管大约 全球三分之一的人口感染结核病，绝大多数感染者坚持 在他们的一生中控制潜在的结核病感染。我们在之前的研究中已经证明，患有 以前治疗过的肺外结核病，没有残留或新感染的迹象，提高了免疫水平 与已治疗的肺结核病患者或已治疗的潜伏感染患者相比，患者的活跃性更强。此外，这些 个体在体外对感染的巨噬细胞产生的细胞因子水平较低。这 这表明易受播散性结核病感染的个体存在潜在的免疫缺陷。我们已经确立了 尖端的分子技术，使我们能够1)直接对病原体-反应性T细胞进行单细胞分类 测序其T细胞受体(TCR)α和β链3)确定T细胞受体α和β 每个T细胞的链和4)重组报告细胞系中的α-β-TCR复合体，以允许 表位特异性的作图。作为概念证明，在本应用程序中，我们将把这些技术应用于 潜伏性结核病患者外周血单核细胞的现有储存库(经治疗和 未经治疗)和以前治疗过的肺结核病或肺外结核病患者。我们假设学位 结核病特异性TCR的多样性可能与个人对潜伏感染的控制程度有关。

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.