ANALYSIS OF A NOVEL SUBSET OF HUMAN CD8+ MEMORY CELLS WITH STEM CELL QUALITIES

具有干细胞特性的新型人类 CD8 记忆细胞亚群的分析

• 批准号: 7832350
• 负责人: STANLEY R. RIDDELL
• 金额: $ 47.97万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-26 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7832350
• 关键词: ABCB1 gene; ATP-Binding Cassette Transporters; Acute; Acute Myelocytic Leukemia; Address; Adult; Age; Antigens; Area; Binding; CD8B1 gene; Cell surface; Cells; Characteristics; Clinical; Clinical Research; Communicable Diseases; Cytomegalovirus; Cytotoxic Chemotherapy; Databases; Development; Disease model; Drug resistance; Effector Cell; Exhibits; Exposure to; Fluorescent Dyes; Frequencies; Gene Expression; Gene Expression Profile; Grant; Granzyme; Hematopoiesis; Hematopoietic stem cells; Heterogeneity; Human; Human Herpesvirus 4; IL2RA gene; Immune response; Immunity; Immunization; Immunologics; In Vitro; Individual; Infection; KLRB1 gene; Latent Virus; Life; Longevity; Lymphoid; Lymphopenia; Maintenance; Malignant - descriptor; Malignant Neoplasms; Memory; Methods; Minor; Modeling; Molecular; Molecular Profiling; Mus; Nature; Organ; Patients; Peripheral; Phenotype; Proliferating; Property; Proteins; Recruitment Activity; Reporter; Resistance; Rhodamine 123; Role; SELL gene; Secondary Immunization; Sorting - Cell Movement; Stem cells; Systems Biology; T memory cell; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Time; Tissues; Transgenic Organisms; Vaccination; Vaccines; Vaccinia; Vaccinia virus; Virus; Virus Diseases; Work; Xenobiotics; chemotherapy; daughter cell; experience; graft vs host disease; improved; in vivo; nonhuman primate; novel; novel strategies; pathogen; programs; public health relevance; reconstitution; resistance mechanism; self-renewal; terminally differentiated effector memory (TEM) T cells

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area 04, Clinical Research and specific Challenge Topic 04-AI-102 - The human immune response to infection and immunization - Profiling via modern immunologic methods and systems biology. The development and maintenance of memory T cells is a hallmark of the host response to acute and persistent viral infections and a major objective of vaccination for infectious diseases and cancer. Analysis of the phenotype and function of memory T cells in both mice and humans has identified two broad subsets of long-lived memory cells termed central memory (TCM), defined by expression of CD45O and CD62L; and effector memory (TEM), defined by expression of CD45RO and the absence of CD62L. Recent work in the applicant's lab has identified a subset of quiescent memory CD8+ T cells in normal individuals that express cell surface markers that enable their separation from the more prevalent TCM and TEM subsets, and includes molecules shared by hematopoietic stem cells. These quiescent memory T cells exhibit higher ATP binding cassette (ABC) transporter activity, rapidly efflux fluorescent dyes and chemotherapy drugs and are resistant to cytotoxic chemotherapy in vitro and in vivo compared with the majority of memory T cells. The high-effluxing T cells have a diverse T cell receptor repertoire, contain T cells specific for persistent and acute viruses, and contribute to repopulation of memory T cells in individuals who have received chemotherapy. Preliminary gene expression arrays performed on sort-purified subpopulations of memory and na¿ve T cells demonstrate that the subsets of memory T cells with high efflux capacity have a distinct transcriptional profile compared with the majority of TM cells, and with TN cells. The studies in this challenge grant will deliver a detailed database on the frequency, functional properties and gene expression profile of these novel subsets of human antigen experienced T cells, and elucidate their development after vaccination. The potential impact of this work relates to improving our understanding of the mechanisms responsible for the durability of human CD8+ T memory in normal individuals and those undergoing chemotherapy or vaccination. The studies have implications for elucidating the nature of protective immunity that might be elicited by vaccines for infectious diseases and cancer, and could provide a marker for the integrity of host immunity to pathogens in aging. The specific aims are: Aim 1. To compare the frequency, transcriptional profile and factors governing maintenance and differentiation of chemotherapy resistant IL-18R?hi CD161hi subsets of human memory CD8+ T cells with IL-18Rlo CD161lo subsets of TCM and TEM, and with TN. The proposed studies will determine the frequency of CMhi and EMhi in normal adult humans (age 18-80), determine the transcriptional profile of these subsets in ten donors, and compare the functional properties of CMhi and EMhi with the respective non-effluxing subsets and with TN. Aim 2. To determine if primary or booster vaccination of humans with vaccinia virus induces a longlived subset of vaccinia virus-specific CD8+ CMhi and EMhi T cells. The time after infection or vaccination when this novel subset of IL18R?hi CD161hi virus-specific T cells might develop has not been defined. The proposed studies will examine the presence of vaccinia virus specific T cells in individuals with remote vaccinia immunization, recent vaccinia boosting and in vaccinia na¿ve individuals receiving a primary vaccination. PUBLIC HEALTH RELEVANCE: The development and maintenance of memory T cells is a hallmark of the host response to acute and persistent viral infections and a major objective of vaccination for infectious diseases and cancer. Progress in defining the qualitative attributes of memory T cells that contribute to their longevity has been much less significant, particularly in humans. Addressing these issues requires a greater understanding of the subsets of T cells that make up the human memory T cell pool and the application of novel approaches to interrogate the cellular and molecular programs that maintain these cells. The studies in this challenge grant will determine the frequency, functional properties and gene expression profile of novel subsets of human memory T cells that have characteristics of long-lived cells, and elucidate their development after vaccination. The potential impact of this work relates to improving our understanding of the mechanisms responsible for the durability of human CD8+ T memory in normal individuals and those undergoing chemotherapy or vaccination.

描述（由申请人提供）：本申请涉及广泛的挑战领域 04、临床研究和特定挑战主题 04-AI-102 - 人体对感染和免疫的免疫反应 - 通过现代免疫学方法和系统生物学进行分析。 记忆T细胞的发育和维持是宿主对急性和持续性病毒感染反应的标志，也是传染病和癌症疫苗接种的主要目标。对小鼠和人类记忆 T 细胞的表型和功能的分析确定了两大类长寿记忆细胞亚群，称为中央记忆 (TCM)，由 CD45O 和 CD62L 的表达定义；和效应记忆 (TEM)，由 CD45RO 的表达和 CD62L 的缺失定义。申请人实验室最近的工作在正常个体中鉴定出了静态记忆 CD8+ T 细胞的子集，这些细胞表达细胞表面标记，使其能够与更普遍的 TCM 和 TEM 子集分离，并且包括造血干细胞共享的分子。与大多数记忆 T 细胞相比，这些静态记忆 T 细胞表现出更高的 ATP 结合盒 (ABC) 转运蛋白活性、快速流出荧光染料和化疗药物，并且在体外和体内对细胞毒性化疗具有抵抗力。高流出 T 细胞具有多样化的 T 细胞受体库，包含针对持久性和急性病毒的特异性 T 细胞，并有助于接受化疗的个体中记忆 T 细胞的重新增殖。对分选纯化的记忆和幼稚 T 细胞亚群进行的初步基因表达阵列表明，与大多数 TM 细胞和 TN 细胞相比，具有高流出能力的记忆 T 细胞亚群具有独特的转录特征。这项挑战资助的研究将提供关于这些新的人类抗原经历 T 细胞亚群的频率、功能特性和基因表达谱的详细数据库，并阐明它们在疫苗接种后的发育。这项工作的潜在影响涉及提高我们对正常个体和接受化疗或疫苗接种的人 CD8+ T 记忆持久性机制的理解。这些研究对于阐明传染病和癌症疫苗可能引发的保护性免疫的本质具有重要意义，并且可以为衰老过程中宿主对病原体免疫的完整性提供标记。具体目标是： 目的 1. 比较人类记忆 CD8+ T 细胞的化疗耐药性 IL-18R?hi CD161hi 亚群与 TCM 和 TEM 的 IL-18Rlo CD161lo 亚群以及 TN 的频率、转录谱和控制维持和分化的因素。拟议的研究将确定正常成年人（18-80 岁）中 CMhi 和 EMhi 的频率，确定十个供体中这些子集的转录谱，并将 CMhi 和 EMhi 与各自的非外排子集和 TN 的功能特性进行比较。目标 2. 确定人类痘苗病毒初次或加强疫苗接种是否会诱导痘苗病毒特异性 CD8+ CMhi 和 EMhi T 细胞的长寿亚群。感染或接种疫苗后，这种新的 IL18R?hi CD161hi 病毒特异性 T 细胞亚群可能发育的时间尚未确定。拟议的研究将检查远程痘苗免疫、近期痘苗加强接种以及接受初次疫苗接种的未接种过痘苗的个体中是否存在痘苗病毒特异性 T 细胞。 公共卫生相关性：记忆 T 细胞的发育和维持是宿主对急性和持续性病毒感染反应的标志，也是传染病和癌症疫苗接种的主要目标。记忆 T 细胞的定性属性有助于其寿命，但在定义这些定性属性方面的进展却不太显着，尤其是在人类中。解决这些问题需要更深入地了解构成人类记忆 T 细胞库的 T 细胞亚群，并应用新方法来探究维持这些细胞的细胞和分子程序。这项挑战资助的研究将确定具有长寿细胞特征的新型人类记忆 T 细胞亚群的频率、功能特性和基因表达谱，并阐明它们在疫苗接种后的发育。 这项工作的潜在影响涉及提高我们对正常个体和接受化疗或疫苗接种的人 CD8+ T 记忆持久性机制的理解。