Genomic Approaches to the T Cell Receptor Repertoire during Aging

衰老过程中 T 细胞受体库的基因组学方法

• 批准号: 8241452
• 负责人: Ning Jenny Jiang
• 金额: $ 2.82万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8241452
• 关键词: Activities of Daily Living; Age; Aging; Base Sequence; Biological Assay; Blood donor; CD8B1 gene; Cell Separation; Cells; Chronic; Clinical; Cytomegalovirus; Cytomegalovirus Infections; Data; Data Analyses; Defect; Development; Diagnostic; Elderly; Epitopes; Frequencies; Gene Expression; Genes; Genomics; Immune; Immune System Diseases; Immune response; Immunodominant Epitopes; Incidence; Individual; Infection; Inflammation; Informatics; Libraries; Ligands; Link; Memory; Methods; Microfluidics; Phenotype; Population; Predisposition; Regulatory T-Lymphocyte; Research; Role; Running; Staining method; Stains; Surface; Surveys; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Technology; Testing; Time; Translating; Virus; base; chemokine; cohort; cytokine; interest; next generation; peripheral blood; response; senescence; tool

DESCRIPTION (provided by applicant): Much research on immune senescence has been focused on T-cells, mainly because a low numbers of naive T-cells in the peripheral blood are the first sign of immune senescence [1]. Furthermore, changes in the T cell receptor (TCR) repertoire diversity have been linked to aging and immune senescence [2]. Aside from the natural reduction in T-cell renewal and TCR repertoire diversity, cytomegalovirus (CMV) infection has a profound influence on subset distribution, phenotype and potentially also on the function of T cells in the elderly [3-5]. However, it is still unclear whether CMV infection is the driver of T cell immune senescence and if so, to what extent. Despite these interesting and suggestive observations, lacking proper tools that enable the systematic study of the rare CMV specific T cells in CMV negative individuals at both the sequence and functional level hinders the quest for an answer to the above questions. Therefore, we propose to develop: 1) a next-generation sequencing based technology to analyze the TCR sequence repertoire; 2) a tetramer staining and isolation method to examine the TCR functional repertoire; and 3) a microfluidic chip-based single cell quantitative PCR (qPCR) technology to dissect functional capabilities of CMV specific T cells. We will apply these technologies to follow and characterize specific T cell populations in older and younger blood donors and correlate this with the timing of CMV seroconversions in both groups. We hypothesize that due to abnormalities in the global and CMV specific T cell repertoires (holes and/or reduced frequency and/or diversity in both TCR sequence and ligand repertoire) and/or defects in the functional capacities of CMV specific precursor T cells, the elderly are predisposed to CMV infection. Furthermore, prolonged period of an inefficient immune response to the virus may result in desensitized immune responses that further drive the development of immune senescence. Our specific aims are: Aim 1. Study global TCR 1 and 2 chain gene diversity difference in young and elderly cohorts using high-throughput sequencing Aim 2. Correlate T cell receptor sequence repertoire with ligand repertoire and phenotype in T cells specific for CMV immunodominant epitopes isolated from young and elderly cohorts Aim 3. Compare functional capabilities of T cells specific for CMV epitopes isolated from young and elderly cohorts PUBLIC HEALTH RELEVANCE: Data generated from this study will help us understand how the TCR repertoire ages and what is the relation between CMV infection and immune senescence. Technologies developed here will be useful to advance clinical diagnostics in CMV infection and immune senescence.

描述(申请人提供)：许多关于免疫衰老的研究一直集中在T细胞上，主要是因为外周血中低数量的初始T细胞是免疫衰老的第一个迹象[1]。此外，T细胞受体(TCR)谱系多样性的变化与衰老和免疫衰老有关[2]。除了T细胞更新和TCR谱系多样性的自然减少外，巨细胞病毒(CMV)感染对老年人的T细胞亚群分布、表型和潜在的T细胞功能也有深远的影响[3-5]。然而，目前尚不清楚CMV感染是否是T细胞免疫衰老的驱动因素，如果是，程度如何。尽管有这些有趣和有启发性的观察结果，但缺乏适当的工具来在序列和功能水平上对CMV阴性个体中罕见的CMV特异性T细胞进行系统研究，阻碍了对上述问题的探索。因此，我们建议发展：1)基于下一代测序的技术来分析TCR序列；2)四聚体染色和分离方法来检测TCR功能；以及3)基于微流控芯片的单细胞定量聚合酶链式反应(QPCR)技术来分析CMV特异性T细胞的功能。我们将应用这些技术来跟踪和表征年龄较大和较年轻的献血者中特定的T细胞群，并将其与两组中CMV血清转换的时间相关联。我们推测，由于全球和CMV特异性T细胞谱系的异常(TCR序列和配基谱系中的空洞和/或频率和/或多样性降低)和/或CMV特异性前体T细胞功能的缺陷，老年人容易受到CMV感染。此外，长时间对病毒的低效免疫反应可能会导致不敏感的免疫反应，从而进一步推动免疫衰老的发展。目的1.利用高通量测序技术研究青年和老年人群中TCR1和TCR2链基因的多样性差异2.将T细胞受体序列与从青年和老年人群中分离的CMV免疫优势表位特异性T细胞的配位库和表型相关联目标3.比较从青年和老年人群中分离的CMV表位特异性T细胞的功能 公共卫生相关性：这项研究产生的数据将帮助我们了解TCR谱系是如何老化的，以及CMV感染和免疫衰老之间的关系。这里开发的技术将有助于提高CMV感染和免疫衰老的临床诊断水平。