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: 8306718
• 负责人: Cameron John Turtle
• 金额: $ 16.98万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8306718
• 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; Radiation therapy; 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; 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

DESCRIPTION (provided by applicant): 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 1CD3 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-3 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.

应聘者描述：应聘者：我的临床和实验室培训重点是发展自己的职业生涯，成为一名在血液学/肿瘤学、造血干细胞移植(HSCT)和人类免疫学方面具有专长的医生/科学家。我在澳大利亚完成了血液学/肿瘤学的临床专业培训，然后获得了博士学位，与Derek Hart一起研究人类树突状细胞(DC)的生物学，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。我和Stanley Riddell一起在弗雷德·哈钦森癌症研究中心(FHCRC)接受博士后培训，重点是研究人类CD8+记忆T细胞，目的是了解记忆T细胞不同亚群的内在特性，这些特性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，2010年被任命为免疫学项目临床研究助理。 职业目标：我的目标是了解人类CD161和CD8+记忆T细胞在正常和动态平衡状态下的调节机制，并研究这些细胞在移植物抗宿主病(GVHD)和自身免疫中的作用。 研究：在最近发表在《免疫学》杂志上的一篇文章中，我调查了急性白血病细胞毒性化疗期间人类T细胞记忆是如何保持的，并确定了一种自我更新的CD8+T细胞亚群，它表达高水平的ABCB1，使它们能够外流化疗药物，并在化疗引起的淋巴细胞减少期间提供持久的记忆T细胞储存库。它们还表达与IL-17的产生相关的基因，包括RORC和CD161，IL-17是一种高度促炎的细胞因子，与许多自身免疫和炎症性疾病的发病机制有关，包括HSCT后的GVHD。 我的工作表明，17型编程的CD161hi亚群包含了相当大的比例(约22%)的人类CD8+记忆T细胞池，并且容纳了整个人类Tc17群体。尽管它们的转录程序是17型，但由于TCR信号通路的调节，只有少数人分泌IL-17或增殖到1CD3单抗。TCR信号通路的调节可以通过提供共刺激或炎症信号来克服，而这些信号的性质决定了CD161hi细胞的命运，导致维持TCR信号调节的17型程序池中的细胞扩张，或者分化为不再受TCR调节抑制的分泌干扰素-3的Tc1样效应细胞。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，可以在允许的炎症环境中释放出来，潜在地导致炎症级联反应的启动。HSCT前条件化放疗诱导的炎症反应可提供必要的启动信号，使CD161中CD8+T细胞失去TCR调节或分化，从而参与GVHD的发病。这项建议的具体目标是： 目的1.研究CD161在中药和透射电子显微镜下CD8+T细胞中TCR信号转导的调控机制。这些研究将定位健康人CD161中CD8+T细胞中TCR信号下调的部位，从而指出炎症性疾病中潜在的调控异常部位。 目的2.研究清髓性异基因HSCT后CD8+T细胞CD161亚群的恢复和功能，并探讨该T细胞亚群是否与GVHD有关。这些研究将确定清髓性异基因HSCT后CD161hi和CD161lo、TEMCD8+细胞的恢复动力学，以及CD161hi CD8+细胞在血液或浸润性组织中的数量是否与急性GVHD有关。然后，我将确定与健康人相比，HSCT患者的TCR信号通路调节是否发生变化，以及HSCT患者CD161hi CD8+细胞中TCR信号调节异常是否与GVHD相关。职业发展和环境：指导的K99阶段将使我能够在我对CD161和CD8+T细胞的初步研究中所观察到的基础上再接再厉。具体地说，在K99阶段，我将发展必要的技能，利用慢病毒载体研究独特调节的CD161hi亚群中的TCR信号，用于基因重建和原代人类T细胞的击倒研究。我将在Riddell博士和专家技术顾问Tewari博士和Randolph-Habecker博士的指导下完成K99研究。在K99期获得的数据和技能将有助于分析CD161hi细胞在GVHD中R00期的调控。在K99/R00阶段开发的技能将为未来的R01应用奠定基础，重点是进一步了解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向独立过渡的理想环境。免疫学计划由四名资深教员组成，他们在计划内外都有卓有成效的合作历史。FHCRC拥有杰出的科学领导力、资源和一流的临床HSCT项目，该项目将允许获取样本，并使HSCT临床医生和科学家能够进行研究合作，并确保我在K99/R00申请中提出的研究成功。