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: 8906718
• 负责人: Ning Jenny Jiang
• 金额: $ 24.15万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8906718
• 关键词: Activities of Daily Living; Age; Aging; Base Sequence; Blood donor; CD8B1 gene; Cell Separation; Cells; Chronic; Clinical; Cytomegalovirus; Cytomegalovirus Infections; Data; Data Analyses; Defect; Development; Diagnostic; Elderly; Epitopes; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genomic approach; High-Throughput Nucleotide Sequencing; 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; deep sequencing; interest; next generation sequencing; peripheral blood; response; senescence; seroconversion; 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.项目摘要/摘要 关于免疫衰老的研究一直集中在T细胞上，主要是因为T细胞数量较少。 外周血液中的幼稚T细胞是免疫衰老的第一个迹象[1]。此外， T细胞受体(TCR)谱系的多样性与衰老和免疫衰老有关[2]。除了 在T细胞更新和TCR谱系多样性的自然减少，巨细胞病毒(CMV)感染具有 对T细胞亚群的分布、表型以及潜在的T细胞功能都有深远的影响 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 如果是这样，在多大程度上。尽管有这些有趣和有启发性的观察，但缺乏适当的工具来实现 CMV阴性个体罕见的CMV特异性T细胞序列和序列的系统研究 职能层面阻碍了对上述问题的寻求答案。因此，我们建议发展：1) 基于下一代测序的分析TCR序列谱的技术；2)四聚体 检测TCR功能谱的染色和分离方法；以及3)基于微流控芯片的 单细胞定量聚合酶链式反应(QPCR)技术用于分析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血液中特定的T细胞群 并将其与两组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(空洞和/或 TCR序列和配基谱系中的频率和/或多样性降低)和/或缺陷 具有CMV特异性前体T细胞功能的老年人易受CMV感染。 此外，长时间对病毒的无效免疫反应可能会导致脱敏。 进一步推动免疫衰老发展的免疫反应。我们的具体目标是： 目的1.研究TCR�和�链基因多样性在青年和老年人群中的差异 高通量测序 目的2.T细胞受体序列谱系与T细胞表型和配基谱系的相关性 从青年和老年人群中分离的CMV免疫优势表位的特异性 目的3.比较青年和成人CMV表位特异性T细胞的功能 老年群体

项目成果

COLT: Constrained Lineage Tree Generation from Sequence Data.

COLT：从序列数据生成约束谱系树。

• DOI: 10.1109/bibm.2016.7822500
• 发表时间: 2016
• 期刊: Proceedings. IEEE International Conference on Bioinformatics and Biomedicine
• 作者: Chen,Keke;Gogu,VenkataSaiAbhishek;Wu,Di;Ning,Jiang
• 通讯作者: Ning,Jiang

Direct measurement of T cell receptor affinity and sequence from naïve antiviral T cells.

• DOI: 10.1126/scitranslmed.aaf1278
• 发表时间: 2016-06-01
• 期刊: Science translational medicine
• 影响因子: 17.1
• 作者: Zhang SQ;Parker P;Ma KY;He C;Shi Q;Cui Z;Williams CM;Wendel BS;Meriwether AI;Salazar MA;Jiang N
• 通讯作者: Jiang N

Immune engineering: from systems immunology to engineering immunity.

免疫工程：从系统免疫学到工程免疫。

• DOI: 10.1016/j.cobme.2017.03.002
• 发表时间: 2017
• 期刊: Current opinion in biomedical engineering
• 影响因子: 3.9
• 作者: Jiang,Ning