Determining Protective Features of Human Memory T-cells to Inform Mycobacterium tuberculosis Vaccine Development

确定人类记忆 T 细胞的保护特征，为结核分枝杆菌疫苗的开发提供信息

基本信息

批准号：
10449464
负责人：
Stephen Matthew Carpenter
金额：
$ 19.33万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-11 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10449464
关键词：
Adolescent Adult Animal Model Antigens Autologous Avidity Benchmarking Bioinformatics Biological Models Bronchoalveolar Lavage Fluid CD4 Positive T Lymphocytes Cell Separation Cell physiology Cells Cellular biology Cessation of life Chronic Clinical Clinical Investigator Award Clinical Research Coculture Techniques Collaborations Communicable Diseases Communities Data Development Disease Environment Flow Cytometry Frequencies Funding Goals HIV Household Human Immune Immunologist Immunology Individual Infection Link Lung Lymphocyte Mentors Mycobacterium tuberculosis Mycobacterium tuberculosis antigens Peptide/MHC Complex Peptides Persons Phenotype Physicians Positioning Attribute Predisposition Preparation Prevention Research Research Personnel Research Priority Severities Specificity System T cell response T memory cell T-Cell Depletion T-Cell Receptor T-Lymphocyte Testing Training Translational Research Tuberculosis Tuberculosis Vaccines Uganda Universities Vaccine Design Vaccines Work antigen-specific T cells chronic infection cohort cytokine effector T cell experience experimental study fighting macrophage medical schools memory CD4 T lymphocyte mouse model prevent programs recruit risk stratification single cell analysis single-cell RNA sequencing skills trafficking transcriptomics vaccine candidate vaccine development

项目摘要

Project Summary / Abstract Tuberculosis (TB), caused by infection with Mycobacterium tuberculosis (Mtb), is a disease that kills 1.4 million people every year. There is no reliable vaccine to prevent TB, yet many latently-infected individuals are protected from progressing to active TB despite heavy Mtb exposure living in an endemic setting. CD4+ T cells are critical for host protection against TB as they interact directly with Mtb-infected cells, secrete cytokines and cytolytic molecules, and recruit or augment other immune cells. However, the candidate and others find that not all T cells specific for Mtb antigens are able to recognize Mtb-infected macrophages, the niche cell for Mtb. A critical unmet need for vaccine development is to define the antigen specificities and functions of T cells that can recognize infected macrophages and prevent progression to active TB. In Aim 1, the candidate uses autologous ex vivo co-culture and T cell antigen receptor (TCR) sequencing to determine the proportion, antigen specificities and functions of memory CD4+ T cells that recognize Mtb-infected macrophages. In Aim 2, the candidate expands on this system to compare the repertoires of memory CD4 T cells that respond to infected macrophages among two groups of individuals who live in a setting endemic for TB yet differ in their susceptibility to active disease. Using single-cell transcriptomics, the candidate compares the functions and TCR repertoires of Mtb- specific memory CD4+ T cells isolated from exposed individuals who do not develop active TB (“stable” latent Mtb infection) vs. individuals who will later progress to active TB (“pre-TB” progressors). Results from this project will define key features of protective memory CD4+ T cells that are linked to the prevention of active TB, providing benchmarks for vaccine development and improvement of TB risk stratification. This 5-year K08 program provides mentoring, training in human immunology, translational research and single-cell transcriptomics for Dr. Stephen Carpenter, a T cell immunologist and infectious disease physician at Case Western Reserve University (CWRU). The institutional environment at CWRU combines an established TB research unit, BSL-3 flow cytometry, cell sorting, single-cell RNA sequencing, and expertise in bioinformatics together with a premier graduate program for scientific interaction and courses. The longstanding Uganda- CWRU Research Collaboration for TB enables clinical and translational work in a TB endemic setting. The candidate is establishing a lab with the long-term goals of understanding the defining features of protective memory T cell responses to Mtb. He has recruited mentors with substantial expertise in translational TB research, T cell biology and single-cell transcriptomics, including Drs. Henry Boom and Mark Cameron at CWRU, and Dr. Sam Behar (UMass Medical School). Completion of this K08 project will transition the candidate into an independent investigator, positioning his research program to use powerful single-cell immune profiling approaches to track and study antigen-specific human memory T cells.

项目摘要 /摘要结核病（TB）是由结核分枝杆菌（MTB）感染引起的，是一种杀死1.4的疾病每年有百万人。没有可靠的疫苗来预防结核病在内在环境中生活在活跃的结核病目的地的活动中。 CD4+ T细胞对于宿主对TB的保护至关重要，因为它们与MTB感染的细胞，秘密细胞因子和细胞溶解分子，并募集或增强其他免疫细胞。但是，候选人和其他人发现没有所有针对MTB抗原的T细胞都能够识别MTB感染的巨噬细胞，即MTB的小裂细胞。一个对疫苗开发的关键未满足需求是定义T细胞的抗原规范和功能公认的感染巨噬细胞并防止进展为活性结核。在AIM 1中，候选人使用自动离体共培养和T细胞抗原受体（TCR）测序以确定比例，抗原规范记忆CD4+ T细胞的功能识别MTB感染的巨噬细胞。在AIM 2中，候选人扩展该系统以比较对感染巨噬细胞响应的记忆CD4 T细胞的库在两组生活中，他们在TB的环境中却有不同的敏感性疾病。使用单细胞转录组学，候选者比较了MTB-的功能和TCR曲目特定的内存CD4+ T细胞从不发展活性TB的暴露个体中分离出来（“稳定”潜在 MTB感染）与后来将进步为主动结核病（“ Pre-TB”进步者）的个人。该项目的结果将定义链接到预防活性结核的受保护内存CD4+ T单元的关键特征用于疫苗开发和改善结核病风险分层的基准。这个为期5年的K08计划提供了人类免疫学，翻译研究和 T细胞免疫学家和传染病医师Stephen Carpenter博士的单细胞转录组学案例西部储备大学（CWRU）。 CWRU的机构环境结合了一个已建立的结核病研究单元，BSL-3流式细胞仪，细胞分选，单细胞RNA测序以及生物信息学方面的专业知识以及一项主要的科学互动和课程研究生课程。长期存在的乌干达 - TB的CWRU研究合作可以在结核病内分子环境中进行临床和翻译工作。这候选人正在建立一个实验室，其长期目标是了解保护的定义特征记忆T细胞对MTB的响应。他招募了具有丰富专业知识的导师，在转化结核病研究方面， T细胞生物学和单细胞转录组学，包括DRS。 CWRU的Henry Boom和Mark Cameron以及博士 Sam Behar（UMass医学院）。该K08项目的完成将使候选人转变为独立研究者，将他的研究计划定位以使用强大的单细胞免疫分析跟踪和研究抗原特异性人体记忆T细胞的方法。