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: 7937922
• 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/7937922
• 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，临床研究和特定的挑战主题04-AI-102 -人类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的反应的标志，也是传染病和癌症疫苗接种的主要目标。对小鼠和人类中记忆T细胞的表型和功能的分析已经鉴定了两个广泛的长寿记忆细胞亚群，称为中央记忆（TCM），由CD 45 O和CD 62 L的表达定义;和效应记忆（TEM），由CD 45 RO的表达和CD 62 L的缺乏定义。申请人实验室的最近工作已经鉴定了正常个体中的静止记忆CD 8 + T细胞的子集，其表达使其能够与更普遍的TCM和TEM子集分离的细胞表面标志物，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，这些静止的记忆T细胞表现出更高的ATP结合盒（ABC）转运体活性，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抗性。高流出性T细胞具有不同的T细胞受体库，含有对持久性和急性病毒特异性的T细胞，并有助于接受化疗的个体中记忆T细胞的再增殖。对记忆和幼稚T细胞的分选纯化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，具有高外排能力的记忆T细胞亚群具有独特的转录谱。这项挑战资助的研究将提供有关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，并阐明它们在接种疫苗后的发展。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CD 8 + T记忆持久性机制的理解。这些研究对于阐明感染性疾病和癌症疫苗可能引起的保护性免疫的性质具有意义，并且可以为衰老中宿主对病原体的免疫完整性提供标记。具体目标是：目标1。比较化疗耐药IL-18 R的频率、转录谱和维持和分化的调控因素？人记忆性CD 8 + T细胞的hi CD 161 hi亚群与TCM和TEM的IL-18 Rlo CD 161 lo亚群以及与TN。所提出的研究将确定CMhi和EMhi在正常成年人（年龄18-80）中的频率，确定这些子集在10个供体中的转录谱，并将CMhi和EMhi的功能特性与相应的非流出子集和TN进行比较。目标二。确定人用牛痘病毒初次或加强接种是否诱导长寿命的牛痘病毒特异性CD 8 + CMhi和EMhi T细胞亚群。感染或接种疫苗后的时间，当这种新的IL 18 R亚群？hi CD 161 hi病毒特异性T细胞的可能发展尚未确定。拟议的研究将检查远程接种牛痘、最近接种牛痘加强的个体以及接受初次疫苗接种的牛痘初治个体中牛痘病毒特异性T细胞的存在。 公共卫生关系：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的反应的标志，也是传染病和癌症疫苗接种的主要目标。在定义记忆T细胞的定性属性方面的进展，特别是在人类中，对记忆T细胞的寿命有贡献，这方面的进展要小得多。解决这些问题需要更好地了解组成人类记忆T细胞库的T细胞亚群，以及应用新方法来询问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人类记忆T细胞新亚群的频率，功能特性和基因表达谱，并阐明它们在疫苗接种后的发展。 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CD 8 + T记忆持久性机制的理解。