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: 8639793
• 负责人: Cameron John Turtle
• 金额: $ 23.41万
• 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/8639793
• 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; 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

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.

抽象的 候选人：我将我的临床和实验室培训集中在发展作为身体/科学家的职业上 具有血液学/肿瘤学，造血干细胞移植（HSCT）和人类方面的专业知识 免疫学。我在澳大利亚完成了血液学/肿瘤学的临床专科培训，然后获得了 研究人类树突细胞生物学（DC）的博士学位，其目的是利用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 斯坦利·里德尔（Stanley Riddell）在弗雷德·哈钦森癌症研究中心（FHCRC）的培训中专注于研究 人类CD8+记忆T细胞的目标，目的是了解不同子集的内在品质 可能预测其在免疫记忆和炎症性疾病中作用的记忆T细胞。 2008年，我开始 担任HSCT服务的主体医师，并于2010年被任命为临床伙伴 免疫学计划的研究。 职业目标：我的目标是了解规范人CD161HI CD8+的机制 记忆T细胞处于正常和扰动的稳态中，并研究这些细胞在移植中的作用 宿主病（GVHD）和自身免疫性。 研究：在最近发表在免疫力的工作中，我研究了人类T细胞记忆如何保持 在急性白血病的细胞毒性化疗期间，并确定了自我更新CD8+ T细胞的子集 表达高水平的ABCB1，使它们能够排出化学疗法药物并提供持久性 化学疗法期间记忆T细胞的储层诱导淋巴细胞减少症。他们也表达基因， 包括与IL-17的产生有关的RORC和CD161，这是一种高度促炎的细胞因子 在许多自身免疫性和炎症性疾病（包括HSCT之后的GVHD）的发病机理中实施。 我的工作表明，17型CD161HI子集包含一个非常大的，并且 以前无法识别的人类CD8+记忆T细胞池的比例（约22％），并拥有整个 人类TC17人口。尽管他们的17型转录计划，但只有少数派分泌IL-17或 由于TCR信号通路的调节，增殖至CD3 mAb。 TCR信号通路调节 可以通过提供共刺激或炎症信号来克服这些信号的性质 CD161HI细胞的命运，导致在维持的17型编程池中的细胞扩展 调节TCR信号传导或分化为不再是分泌的TC1样效应细胞的IFN- 受TCR调节的约束。这些数据表明CD161HI CD8+子集是一个非常大的 可以在宽松的炎症中释放的17型型记忆单元的储层 环境，有可能导致炎症级联反应。引起的炎症 在HSCT之前调节化学疗法，可以提供必要的启动信号，导致损失 CD161HI CD8+ T细胞中的TCR调节或分化，使其有助于其发病机理 GVHD。该提案的具体目的是： 目的1。表征调节CD161HI TCM和TEM CD8+中TCR信号传递的机制 T细胞。这些研究将在CD161HI CD8+ T细胞中定位TCR信号传导下调的位点 健康的个体，从而表明炎症性疾病失调的潜在部位。 AIM 2。表征骨髓性后CD161HI CD8+ T细胞的恢复和功能 同种异体HSCT，并确定该T细胞子集是否在GVHD中暗示。这些研究将确定 骨髓性同种异体HSCT后，CD161HI和CD161LO TCM和TEM CD8+细胞的回收动力学， 如果血液或浸润组织中的CD161HI CD8+细胞数与急性GVHD有关。然后我会 确定与健康个体相比，HSCT患者的TCR信号通路调节是否改变了 如果来自HSCT患者的CD161HI CD8+细胞中TCR信号的失调与GVHD相关。 职业发展和环境：事物的K99阶段将使我能够以观察为基础 在我对CD161HI CD8+ T细胞的初步研究中进行的。具体来说，在K99阶段，我将发展技能 使用慢病毒载体在唯一调节的CD161HI子集中研究TCR信号是必要的 基因重建和对原代人T细胞的敲低研究。我将通过 Riddell博士和专家技术顾问的指导。 Tewari和Randolph-Habecker。数据 获得K99阶段的技能将有助于分析R00中GVHD中CD161HI细胞的调节 阶段。 K99/R00阶段开发的技能将为以后的R01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的科学和临床环境非常适合职业发展以及过渡到 独立。免疫学计划已完成四名高级教职员工，他们有历史 计划内外的产品协作。 FHCRC具有出色的科学领导， 资源和精湛的临床HSCT计划，该计划将允许获取样品并启用 与HSCT临床医生和科学家的研究合作，并确保提出的研究成功 我的K99/R00应用程序。