Longitudinal Analysis of Immune Signatures (IMS) of M. tuberculosis-specific T cells

项目摘要：项目2 肺结核是由结核分枝杆菌(Mtb)感染引起的，是一种主要的 世界范围内单一感染源致死的原因。在这里，我们将描述免疫特征(IM) 不同结核分枝杆菌感染状态和卡介苗接种人群中结核分枝杆菌抗原特异性CD4T细胞的研究 具体地说，我们将利用我们建立的Mtb多肽表位库和检测系统来识别抗原- 特异性CD4T细胞及其T细胞受体(TCRs)。IMS将进一步在细胞学和 抗原刺激后和体外进行单细胞rna-seq检测。对于目标1，我们将招募一批个人 患有活动性肺结核(ATB)，特征是无控制的有症状的结核分枝杆菌感染，谁将接受 抗结核治疗。我们将在治疗过程中纵向跟踪IMS，并确定标志物 预测治疗结果。对于目标2，我们将招收患有潜在结核病感染(LTBI)的个人，这是 具有对结核分枝杆菌的免疫反应性，但没有疾病症状(即结核分枝杆菌反应)。LTBI是 异质性，由成功清除感染并表现出 记忆免疫反应，以及其他亚临床感染，发展为活动性增加的风险 疾病。目前还没有可用的测试来区分亚临床感染和明确的感染。我们将决定 经纵向预防治疗的LTBI的IMS，并决定谁对治疗反应最大 类似于ATB，我们认为它最有可能患有亚临床疾病。对于LTBI的单独队列， 我们将利用一种独特的人类挑战模型，从 并使用它们来评估血液中的IMS与来自同一肺的IMS之间的重叠和差异 一组个体。对于目标3，我们将获取成人在接种卡介苗(再)疫苗前和之后的样本，以 描述疫苗诱导的IMS的特征。未感染结核分枝杆菌的成人复种被证明是有保护作用的 因此，将这一IMS与LTBI(控制感染)和ATB(非控制感染)中的IMS进行比较 可以揭示疫苗诱导免疫的保护性成分。

Project Summary: Project 2 Pulmonary tuberculosis (TB) is caused by infection with Mycobacterium tuberculosis (Mtb) and is a leading cause of death from a single infectious agent worldwide. Here we will characterize immune signatures (IMS) of Mtb antigen-specific CD4 T cells in individuals with different states of Mtb infection and BCG vaccination. Specifically, we will utilize our established Mtb peptide epitope pools and assay systems to identify antigen- specific CD4 T cells and their T cell receptors (TCRs). The IMS will be further characterized in cytometry and single-cell RNA-seq assays after antigen stimulation and ex vivo. For Aim 1, we will enroll cohorts of individuals with active tuberculosis (ATB), characterized by uncontrolled symptomatic Mtb infection, who will be receiving anti-TB therapy. We will longitudinally track the IMS over the course of treatment, and identify markers predictive of treatment outcomes. For Aim 2, we will enroll individuals with ‘latent’ TB infection (LTBI), which is characterized by immune reactivity to Mtb but no symptoms of disease (i.e. Mtb reactive). LTBI is heterogeneous, consisting both of individuals who have successfully cleared their infection and display a memory immune response, as well as others with subclinical infection at increased risk of developing active disease. There is currently no test available to distinguish subclinical from cleared infection. We will determine the IMS in prophylactically-treated LTBI longitudinally, and determine who displays a treatment response most similar to ATB, which we consider most likely to have had subclinical disease. For a separate cohort of LTBI, we will utilize a unique human challenge model to obtain samples with antigen-specific resident T cells from the lung and use them to evaluate overlap and differences between the IMS in the blood vs. lung from the same set of individuals. For Aim 3, we will obtain samples taken pre- and post-BCG (re)vaccination in adults to characterize the vaccine induced IMS. Revaccination of Mtb uninfected adults has been shown to be protective from disease, thus, comparing this IMS with that in LTBI (controlled infection) vs. ATB (uncontrolled infection) can reveal protective components of vaccine induced immunity.