Evolution & Maintenance of Memory CD8 T Cells

进化

• 批准号: 7450940
• 负责人: Katherine F Luzuriaga
• 金额: $ 161.89万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-24 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7450940
• 关键词: Evolution; Maintenance; Memory; CD8; Cells

DESCRIPTION (provided by applicant): This Program Project proposes in-depth studies of the development of CD8+ T lymphocyte responses to persistent (EBV) and non-persistent (vaccinia, yellow fever) virus infections with particular emphasis on understanding induction and long-term maintenance of antigen-specific and cross-reactive CD8+ T cells. The overall goals are: 1) To use models of persistent (EBV) and non-persistent (VV or YFV) viral infections to characterize the lineage relationship between effector and memory responses in humans and the factors that influence evolution of antigen-specific CD8+ T cell responses into the memory CD8+ T cell repertoire; 2) To characterize how cross-reactivity influences the evolution of the antigen-specific CD8+ T cell repertoire and understand the structural and functional properties of the T cell receptor that contribute to recognition of antigen-specific or cross-reactive ligands; 3) To correlate antigen-specific and cross-reactive CD8+ T cell frequencies and functional properties with control of viral replication or manifestations of disease. Project #1 will use the Epstein Barr model of persistent viral infection to characterize the lineage relationship between effector and memory responses and the factors that influence evolution of antigen-specific CD8+ T cell responses into the memory CD8+ T cell repertoire. Project #2 will test the hypothesis that the quantity, quality, and avidity of an epitope-specific response to acute viral infection are dramatically altered by acute heterologous virus infections and may alter disease outcome. It will also test the hypothesis that, because of prior antigenic exposure, adults may have higher frequencies of cross-reactive CD8+ T cells than children. TCR alpha and beta chain spectratyping and sequencing will allow improved understanding of the structural bases for the recognition of antigen-specific or cross-reactive ligands. Project #3 will study vaccinia virus or yellow fever virus vaccine recipients to characterize the lineage relationship between effector and memory responses in humans and the factors that influence evolution of antigen-specific CD8+ T cell responses into the memory CD8+ T cell repertoire. This project will also test the hypothesis that VV or YFV infection will reactivate cross-reactive CD8+ T cells. The proposed work will benefit from a highly interactive and collaborative group of productive investigators. Four core facilities will facilitate the work of Project Investigators. The Administrative and Clinical Core (Core A) will coordinate the Projects; manage fiscal affairs; be responsible for the collection and processing of clinical specimens from research subjects with AIM or in the latent phase of EBV infection; provide EBV serology and molecular HLA Class I and II typing of research subjects. The Tetramer Core (Core B) will provide HLA Class l/peptide tetramers to all Project Investigators. The Flow Cytometry (Core C) will provide FACS-based cell sorting and characterization of virus-specific or cross-reactive CD8+ T cell responses. A TCR Core (Core D) will perform spectratyping, TCR subcloning and sequencing, maintain a TCR repertoire database, and facilitate TCR data analyses. The proposed studies will help us to understand the timing of and factors that control the evolution of human antiviral CD8+ T cell responses from the acute effector phase into long-term memory. These studies will also help us to understand variability in human responses to viral infections and how antigen-specific or cross-reactive CD8+ T cells contribute to either viral control or disease. Understanding the mechanisms operative in the evolution of virus-specific CD8+ T cell responses should contribute to the development of new or improved viral vaccines and the development of novel therapies for autoimmune diseases or virus associated malignancies. PROJECT 1: "Evolution and Maintenance of EBV-Specific Memory CD8+ T Cells" (Luzuriaga, K.) PROJECT 1 DESCRIPTION (provided by applicant): Work outlined in this proposal addresses the hypotheses that virus-specific CD8+ T cell repertoires change quantitatively and qualitatively over time following acute infection, and that differences in virus-specific CD8+ T cell repertoires may influence the control of viral replication or disease pathogenesis. Changes in the EBV-specific CD8+ T cell repertoire over time may be based on properties of EBV-specific CD8+ T cell clones (specificity, T cell receptor usage, cytokine receptors, and expression of survival factors) and ongoing antigen exposure. Collaborative work with Drs. Selin and Welsh (Project 3) will test the hypothesis that age-related differences in the evolution of cross-reactive CD8+ T lymphocyte responses will be observed and may be influenced by prior (or lack of prior) exposure to other viral pathogens. To address these hypotheses, we will investigate the evolution of CD8+ T lymphocyte responses during the early (induction) and long-term (maintenance) phases of the immune response in infants, children, adolescents, and adults. The following Specific Aims will be addressed: 1) To define the relationship between the virus-specific T cell receptor repertoires at serial time points from the acute through the memory phases of the T cell response; 2) To define the relationship between the expression of cytokine receptors or survival factors during the acute phase of the T cell response and T cell clonal persistence into the memory pool; 3) To define the relationship between the functional profiles of virus-specific T cells during the acute phase of the T cell response and their maintenance of proliferative and cytolytic capacity during the long-term memory phase; and 4) To correlate EBV-specific CD8+ T cell frequencies and functional properties over time with blood viral load, pharyngeal shedding, and disease severity. This project directly supports the overall objective of the Program Project by characterizing the induction, evolution, and maintenance of virus-specific CD8+ T lymphocyte responses to a persistent viral infection. Our investigation of memory CD8+ T lymphocyte responses resulting from a persistent viral infection with repeated but changing exposure to viral antigens will complement Drs. Rothman's and Ennis' investigation of memory CD8+ T lymphocyte responses resulting from receipt of yellow fever or vaccinia vaccines in Project 3. Altogether, these studies will help us to understand the timing of and factors that control the evolution of human antiviral CD8+ T cell responses from the acute effector phase into long-term memory. These studies will also help us to understand how antigen-specific or cross-reactive CD8+ T cells may contribute to either viral control or disease. Ultimately, we hope that they will help to inform the development of antiviral vaccines that afford long-term protection against infection or disease, while minimizing the potential adverse effects of these vaccines.

描述（由申请人提供）：该计划项目提出了对CD8+ T淋巴细胞对持久性（EBV）和非持久性（疫苗，黄热病）病毒感染的反应的深入研究，尤其强调了对抗原特异性和交叉毒性CD8+ T细胞的理解诱导和长期维持。总体目标是：1）使用持久性（EBV）和非持久性（VV或YFV）病毒感染的模型来表征人类效应子与记忆反应之间的谱系关系以及影响抗原特异性CD8+ T细胞反应在存储器中的演化的因素； 2）表征交叉反应性如何影响抗原特异性CD8+ T细胞库的演变，并了解T细胞受体的结构和功能特性，从而有助于识别抗原特异性或交叉反应性配体； 3）将抗原特异性和交叉反应的CD8+ T细胞频率以及功能特性与病毒复制或疾病的表现相关联。项目＃1将使用持续病毒感染的爱泼斯坦BARR模型来表征效应子与记忆反应之间的谱系关系以及影响抗原特异性CD8+ T细胞反应在存储器CD8+ T细胞库中的进化的因素。项目＃2将检验以下假设：表位特异性反应对急性病毒感染的数量，质量和亲和力大大改变了急性异源性病毒感染，并可能改变疾病结果。它还将检验以下假设：由于先前的抗原暴露，成年人的交叉反应性CD8+ T细胞的频率可能高于儿童。 TCRα和β链谱分型和测序将可以提高对抗原特异性或交叉反应性配体识别结构碱基的理解。项目＃3将研究疫苗病毒或黄热病病毒疫苗接受者，以表征人类效应子与记忆反应之间的谱系关系以及影响抗原特异性CD8+ T细胞反应在记忆CD8+ T细胞库中的进化的因素。该项目还将检验以下假设：VV或YFV感染将重新激活交叉反应性CD8+ T细胞。拟议的工作将受益于高度互动和协作的生产研究人员。四个核心设施将促进项目调查人员的工作。行政和临床核心（核心A）将协调项目；管理财政事务；负责从AIM或EBV感染潜在的研究对象收集和处理临床标本；提供研究对象的EBV血清学和分子HLA I类和II类。四聚体核心（核心B）将向所有项目研究人员提供HLA类L/肽四聚体。流式细胞仪（CORE C）将提供基于FACS的细胞分选和病毒特异性或交叉反应性CD8+ T细胞反应的表征。 TCR核心（核心D）将执行频谱型，TCR亚克隆和测序，维持TCR库库数据库并促进TCR数据分析。拟议的研究将帮助我们了解控制人类抗病毒药CD8+ T细胞反应从急性效应阶段演变为长期记忆的因素的时间和因素。这些研究还将帮助我们了解人类对病毒感染的反应的变异性以及抗原特异性或交叉反应性CD8+ T细胞如何有助于病毒控制或疾病。了解病毒特异性CD8+ T细胞反应进化中的机制应有助于开发新的或改善的病毒疫苗，并开发用于自身免疫性疾病或病毒相关的恶性肿瘤的新型疗法。 项目1：“ EBV特异性内存CD8+ T细胞的进化和维护”（Luzuriaga，K。） 项目1描述（由申请人提供）：该提案中概述的工作解决了病毒特异性CD8+ T细胞库的假设，急性感染后随着时间的推移会随着时间的推移而定性地发生变化，并且病毒特异性CD8+ T细胞的差异可能会影响病毒复制或疾病病原体生理基础的病毒复制的控制。随着时间的推移，EBV特异性CD8+ T细胞库的变化可能基于EBV特异性CD8+ T细胞克隆的特性（特异性，T细胞受体使用，细胞因子受体和存活因子的表达）和持续的抗原暴露。与博士的合作工作。 Selin和Welsh（项目3）将检验以下假设：将观察到交叉反应性CD8+ T淋巴细胞反应的年龄相关差异，并且可能会受到先验（或缺乏先验）其他病毒病原体的影响。为了解决这些假设，我们将研究婴儿，儿童，青少年和成人的早期（诱导）和长期（诱导）和长期（维持）阶段CD8+ T淋巴细胞反应的演变。将解决以下特定目的：1）在急性通过T细胞反应的记忆阶段的串行时间点处定义病毒特异性T细胞受体库之间的关系； 2）定义在T细胞反应的急性相和T细胞克隆持久性进入记忆池的急性期中细胞因子受体表达之间的关系； 3）定义在T细胞反应的急性阶段病毒特异性T细胞的功能谱之间的关系，并在长期记忆阶段维持增生和溶液能力的维持； 4）将EBV特异性的CD8+ T细胞频率和功能特性与血液病毒载荷，咽部脱落和疾病严重程度相关联。该项目通过表征对持续病毒感染的病毒特异性CD8+ T淋巴细胞反应的诱导，演变和维护来直接支持该计划项目的整体目标。我们对持续性病毒感染产生的记忆CD8+ T淋巴细胞反应的研究将与病毒抗原的反复接触而变化。罗斯曼（Rothman）和恩尼斯（Ennis）对项目3中的黄热病或疫苗接种导致的记忆CD8+ T淋巴细胞反应的研究。总的来说，这些研究将有助于我们理解人类抗病毒CD8+ T细胞反应的进化，从急性效应阶段进入长期记忆。这些研究还将帮助我们了解抗原特异性或交叉反应性CD8+ T细胞如何有助于病毒控制或疾病。最终，我们希望它们将有助于告知抗病毒疫苗的开发，这些抗病毒疫苗可长期保护感染或疾病，同时最大程度地减少这些疫苗的潜在不利影响。