Quiescent Tc17 programmed CD8 T cells and their role in graft versus host disease

静态 Tc17 编程 CD8 T 细胞及其在移植物抗宿主疾病中的作用

• 批准号: 8685185
• 负责人: Cameron John Turtle
• 金额: $ 24.15万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-18 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8685185
• 关键词: ABCB1 gene; Acute; Acute Graft Versus Host Disease; Acute leukemia; Address; Allogenic; Australia; Autoimmune Diseases; Autoimmune Process; Autoimmunity; B lymphoid malignancy; Bacteria; Blood; CD3 Antigens; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Count; Cells; Clinical; Clinical Research; Collaborations; Communicable Diseases; Complication; Cytotoxic Chemotherapy; Cytotoxic agent; Data; Dendritic Cells; Disease; Doctor of Philosophy; Down-Regulation; Effector Cell; Ensure; Environment; Exhibits; Faculty; Flow Cytometry; Fluorescent Dyes; Foundations; Fred Hutchinson Cancer Research Center; Frequencies; Future; Gene Expression Profiling; Genes; Goals; Hematology; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Homeostasis; Homing; Human; Human Biology; Immune; Immune response; Immunity; Immunology; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Interferons; Interleukin-17; KLRB1 gene; Kinetics; Laboratories; Leadership; Lentivirus Vector; Lymphopenia; Maintenance; Malignant Neoplasms; Mediating; Memory; Mentors; Minority; Mus; Nature; Oncogenic Viruses; Pathogenesis; Pathologic; Pathway Analysis; Patients; Pharmaceutical Preparations; Phase; Physicians; Population; Production; Proliferating; Publishing; Receptor Signaling; Recording of previous events; Recovery; Recovery of Function; Recruitment Activity; Regulation; Research; Research Personnel; Resistance; Resources; Rhodamine 123; Role; SELL gene; Sampling; Scientist; Services; Signal Transduction; Site; Stem cell transplant; Stem cells; T Cell Receptor Signaling Pathway; T cell response; T memory cell; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Tissues; Training; Vaccination; Virus; Work; anergy; base; career; career development; chemoradiation; chemotherapy; conditioning; cytokine; fungus; graft vs host disease; immune activation; insight; knock-down; member; novel; oncology; pathogen; post-doctoral training; programs; reconstitution; response; self-renewal; skills; success; therapy development

ABSTRACT Candidate: I have focused my clinical and laboratory training on developing a career as a physician/scientist with expertise in hematology/oncology, hematopoietic stem cell transplantation (HSCT) and human immunology. I completed clinical subspecialty training in hematology/oncology in Australia, and then obtained a PhD studying the biology of human dendritic cells (DCs) with Derek Hart with the goal of utilizing DCs to generate virus- and tumor-specific CD8+ T cell responses in patients with B cell malignancies. My postdoctoral training at the Fred Hutchinson Cancer Research Center (FHCRC) with Stanley Riddell has focused on studies of human CD8+ memory T cells, with the goal of understanding intrinsic qualities of different subsets of memory T cells that might predict their role in immune memory and inflammatory disease. In 2008, I began serving as an Attending Physician on the HSCT service, and in 2010 was appointed an Associate in Clinical Research in the Program in Immunology. Career Objectives: My objectives are to understand the mechanisms that regulate human CD161hi CD8+ memory T cells in normal and perturbed homeostasis and to investigate a role for these cells in graft versus host disease (GVHD) and autoimmunity. Research: In work recently published in Immunity, I investigated how human T cell memory is maintained during cytotoxic chemotherapy for acute leukemia, and identified a subset of self-renewing CD8+ T cells that expresses high levels of ABCB1, enabling them to efflux chemotherapy drugs and provide a persistent reservoir of memory T cells during chemotherapy induced lymphocytopenia. They also express genes, including RORC, and CD161 that are associated with production of IL-17, a highly pro-inflammatory cytokine implicated in the pathogenesis of many autoimmune and inflammatory diseases, including GVHD after HSCT. My work has revealed that the type 17-programmed CD161hi subset comprises a remarkably large, and previously unrecognized, proportion (~22%) of the human CD8+ memory T cell pool, and harbors the entire human Tc17 population. Despite their type 17- transcriptional program, only a minority secretes IL-17 or proliferates to ¿CD3 mAb due to regulation of the TCR signaling pathway. TCR signaling pathway regulation can be overcome by provision of costimulation or inflammatory signals, and the nature of those signals dictates the fate of CD161hi cells, resulting in either expansion of cells in the type 17 programmed pool that maintain regulation of TCR signaling or differentiation into IFN-¿ secreting Tc1-like effector cells that are no longer restrained by TCR regulation. These data suggest that the CD161hi CD8+ subset is an extraordinarily large reservoir of type 17-programmed memory cells that can be unleashed in a permissive inflammatory environment, potentially resulting in initiation of an inflammatory cascade. Inflammation induced by conditioning chemo-radiotherapy prior to HSCT could provide the necessary initiating signals that cause loss of TCR regulation or differentiation in CD161hi CD8+ T cells and allow them to contribute to the pathogenesis of GVHD. The specific aims of this proposal are: Aim 1. To characterize mechanisms regulating TCR signal transduction in CD161hi TCM and TEM CD8+ T cells. These studies will localize the sites of downregulation of TCR signaling in CD161hi CD8+ T cells in healthy individuals, thereby indicating potential sites of dysregulation in inflammatory diseases. Aim 2. To characterize the recovery and function of CD161hi CD8+ T cells after myeloablative allogeneic HSCT, and determine if this T cell subset is implicated in GVHD. These studies will determine the kinetics of recovery of CD161hi and CD161lo TCM and TEM CD8+ cells after myeloablative allogeneic HSCT, and if CD161hi CD8+ cell numbers in blood or infiltrated tissue are associated with acute GVHD. I will then establish if TCR signaling pathway regulation is altered in HSCT patients compared to healthy individuals and if dysregulation of TCR signaling in CD161hi CD8+ cells from HSCT patients is correlated with GVHD. Career Development and Environment: The mentored K99 phase will allow me to build on the observations made in my preliminary studies of CD161hi CD8+ T cells. Specifically, during the K99 phase I will develop skills that are necessary to study TCR signaling in the uniquely regulated CD161hi subset using lentiviral vectors for gene reconstitution and knock-down studies in primary human T cells. I will complete the K99 studies with guidance from Dr. Riddell and the expert technical consultants, Drs. Tewari and Randolph-Habecker. The data and skills acquired the K99 phase will facilitate analysis of the regulation of CD161hi cells in GVHD in the R00 phase. The skills developed in the K99/R00 phase will lay the foundation for a future R01 application focused on further understanding the role of CD161hi T cells in human inflammatory responses, including autoimmunity. The scientific and clinical environment at FHCRC is ideal for career development and the transition to independence. The Program in Immunology is comprised of four Senior Faculty members, who have a history of productive collaboration within and outside the program. The FHCRC has outstanding scientific leadership, resources, and a superb clinical HSCT program, which will allow the acquisition of samples and enable research collaborations with HSCT clinicians and scientists, and ensure success of the research proposed in my K99/R00 application.

摘要 应聘者：我的临床和实验室培训主要集中在发展医生/科学家的职业生涯上 在血液学/肿瘤学、造血干细胞移植(HSCT)和人类 免疫学。我在澳大利亚完成了血液学/肿瘤学的临床分专业培训，然后获得了 与Derek Hart一起研究人类树突状细胞(DC)生物学的博士学位，目标是利用DC 在B细胞恶性肿瘤患者中产生病毒和肿瘤特异性CD8+T细胞反应。我的博士后 在弗雷德·哈钦森癌症研究中心(FHCRC)与斯坦利·里德尔一起接受培训的重点是研究 人类CD8+记忆T细胞，目的是了解不同亚群的固有性质 记忆T细胞可能预测它们在免疫记忆和炎症性疾病中的作用。2008年，我开始 担任HSCT服务的主治医生，并于2010年被任命为临床助理医生 免疫学项目中的研究。 职业目标：我的目标是了解人类CD161/CD8+的调节机制 记忆T细胞在正常和动态平衡紊乱中的作用以及这些细胞在移植物抗宿主病中的作用 宿主病(GVHD)和自身免疫。 研究：在最近发表在《免疫》杂志上的一篇文章中，我研究了人类T细胞记忆是如何维持的 在急性白血病的细胞毒性化疗期间，发现了一组自我更新的CD8+T细胞 表达高水平的ABCB1，使它们能够外排化疗药物并提供持久的 化疗期间记忆T细胞储存库引起的淋巴细胞减少。它们也表达基因， 包括与高度促炎细胞因子IL-17的产生有关的RORC和CD161 与许多自身免疫性和炎症性疾病的发病机制有关，包括移植后的移植物抗宿主病。 我的工作揭示了17型编程的CD161hi亚集包含一个非常大的，并且 以前未被认识到的，比例(~22%)的人CD8+记忆T细胞池，并包含整个 人类Tc17群体。尽管他们的17型转录程序，只有少数分泌IL-17或 由于TCR信号通路的调节而增殖为CD3单抗。TCR信号通路调控 可以通过提供共刺激或炎症信号来克服，这些信号的性质决定了 CD161hi细胞的命运，导致维持在17型程序化池中的细胞扩张 TCR信号的调节或向不再分泌Tc1样效应细胞的干扰素-β的分化 受到TCR法规的限制。这些数据表明CD161hi CD8+亚群是一个非常大的 17型编程记忆细胞的储存库，可在允许的炎症反应中释放 环境，有可能导致炎症级联反应。引起炎症的原因是 造血干细胞移植前的条件化放化疗可以提供必要的启动信号，从而导致 TCR对CD8+T细胞CD161的调节或分化，并使其在慢性粒细胞白血病的发病机制中起作用 GVHD。这项建议的具体目标是： 目的1.研究中药CD161和透射电子显微镜CD8+中TCR信号转导的调控机制 T细胞。这些研究将定位CD161和CD8+T细胞中TCR信号下调的部位 健康的个体，从而指出炎症性疾病中潜在的调节失调部位。 目的2.研究清髓后CD8+T细胞CD161的恢复和功能 异基因HSCT，并确定这个T细胞亚群是否与GVHD有关。这些研究将确定 清髓性异基因造血干细胞移植后CD161hi和CD161lo及TEMCD8+细胞恢复的动力学研究 CD161hi CD8+细胞在血液或浸润性组织中的数量与急性GVHD相关。那我会的 确定与健康人相比，HSCT患者的TCR信号通路调节是否发生改变 HSCT患者CD161hi CD8+细胞TCR信号异常与GVHD相关。 职业发展和环境：指导K99阶段将使我能够在观察的基础上再接再厉 这是我对CD8+T细胞中CD161的初步研究。具体地说，在K99阶段，我将发展技能 对于使用慢病毒载体研究独特调节的CD161hi亚群中的TCR信号是必要的 原代人类T细胞的基因重组和基因敲除研究。我将完成K99研究 Riddell博士和专家技术顾问Tewari博士和Randolph-Habecker博士的指导。数据 在K99期获得的技能将有助于分析R00中CD161hi细胞在GVHD中的调节 相位。在K99/R00阶段培养的技能将为未来专注于R01的应用程序奠定基础 进一步了解CD161hi T细胞在包括自身免疫在内的人类炎症反应中的作用。 FHCRC的科学和临床环境是职业发展和过渡到 独立。免疫学项目由四名资深教员组成，他们都有 计划内外的富有成效的协作。FHCRC拥有杰出的科学领导力， 资源，以及一流的临床HSCT计划，该计划将允许获取样本并使 与造血干细胞移植临床医生和科学家的研究合作，并确保在 我的K99/R00应用程序。