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: 8334072
• 负责人: Ning Jenny Jiang
• 金额: $ 13.32万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 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 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8334072
• 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; abstracting; base; chemokine; cohort; cytokine; interest; next generation; peripheral blood; response; senescence; tool

7. Project Summary/Abstract 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 ¿ and ¿ 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

7.项目总结/摘要 许多关于免疫衰老的研究都集中在T细胞上，主要是因为T细胞数量很少。 外周血中的幼稚T细胞是免疫衰老的第一个迹象[1]。此外， T细胞受体（TCR）库多样性与衰老和免疫衰老有关[2]。除了 由于T细胞更新和TCR库多样性的自然减少，巨细胞病毒（CMV）感染具有 对亚群分布、表型和潜在的T细胞功能的深远影响 老年人[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，缺乏适当的工具， 系统研究CMV阴性个体中罕见的CMV特异性T细胞， 功能层面的障碍阻碍了对上述问题的回答。因此，我们建议发展：1） 分析TCR序列库的基于下一代测序的技术; 2）四聚体 染色和分离方法，以检查TCR功能库;和3）基于微流控芯片的 单细胞定量PCR（qPCR）技术来剖析CMV特异性T细胞的功能能力。我们 将应用这些技术来跟踪和表征老年人和年轻人血液中的特定T细胞群 并将其与两组中CMV血清转换的时间相关联。 我们假设，由于总体和CMV特异性T细胞库的异常（孔和/或 TCR序列和配体库的频率和/或多样性降低）和/或TCR序列和配体库的缺陷。 由于CMV特异性前体T细胞的功能能力，老年人易受CMV感染。 此外，长时间对病毒的无效免疫应答可能导致脱敏。 免疫反应，进一步推动免疫衰老的发展。我们的具体目标是： 目标1.研究年轻和老年队列中TCR和TCR链基因多样性的差异， 高通量测序 目标2.将T细胞受体序列库与T细胞中的配体库和表型相关联 对分离自年轻和老年人群的CMV免疫显性表位具有特异性 目标3.比较从年轻人和成年人中分离的CMV表位特异性T细胞的功能能力， 老年队列