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: 8185834
• 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 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8185834
• 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. PUBLIC HEALTH RELEVANCE: The applicant has identified a new way to isolate and study a large proportion of the immune memory cells that provide protection after vaccination or infection. The memory T cells that the applicant has identified in humans are resistant to chemotherapy, which allows them to survive in chemotherapy-treated patients and contribute to the reconstitution of immune memory to viruses after recovery from chemotherapy. The chemotherapy- resistant cells have the potential to cause profound inflammation, but do not do so in healthy individuals because of stringent regulation. Studies to identify mechanisms that cause a loss of this regulation might provide information on the causes and treatment of graft versus host disease (GVHD), a major complication of blood stem cell transplantation, and other inflammatory or autoimmune diseases. Additional research might allow the development of treatments to increase immunity to infections and cancer after chemotherapy or blood stem cell transplantation.

描述（由申请人提供）：候选人：我将临床和实验室培训重点放在发展为血液学/肿瘤学，造血干细胞移植（HSCT）和人类免疫学专业知识的医师/科学家的职业上。我在澳大利亚完成了血液学/肿瘤学领域的临床专科培训，然后获得了研究人类树突状细胞（DCS）的生物学博士学位，其目的是利用DCS生成病毒和肿瘤特异性CD8+ T细胞的CD8+ T细胞反应。我在斯坦利·里德尔（Stanley Riddell）的弗雷德·哈钦森癌症研究中心（FHCRC）在弗雷德·哈钦森癌症研究中心（FHCRC）的博士后培训专注于人类CD8+记忆T细胞的研究，其目的是了解不同记忆T细胞的内在品质，这可能预测了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服务的医师，并于2010年被任命为免疫学计划临床研究的助理。 职业目标：我的目标是了解调节正常和扰动稳态中人类CD161HI CD8+记忆T细胞的机制，并研究这些细胞在移植物与宿主疾病（GVHD）和自身免疫性中的作用。 研究：在最近发表的免疫性工作中，我研究了在急性白血病的细胞毒性化学疗法期间如何维持人T细胞记忆，并确定了自我更新的CD8+ T细胞的子集，以表达高水平的ABCB1，使它们能够使它们能够在化学疗法中均能储存在化学疗法的过程中，并提供了在化学治疗过程中的含量。它们还表达了与IL-17的产生有关的基因，包括RORC和CD161，这是一种高度促炎性细胞因子，涉及许多自身免疫性和炎症性疾病的发病机理，包括HSCT后的GVHD。 我的工作表明，17型CD161HI子集包含人类CD8+记忆T细胞池的一个非常大的，以前未被认可的比例（约22％），并拥有整个人类TC17人群。尽管它们的17型转录程序，但由于TCR信号通路的调节，只有少数族裔分泌IL-17或增殖至1CD3 mAb。 TCR信号通路路径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，从而导致在17型编程池中的细胞扩展，以维持TCR信号的调控，或者将TCR信号的调节或分化为IFN-3的IFN-3分离为TC1样细胞不再受到TCR调节的限制。这些数据表明，CD161HI CD8+子集是可以在允许性的炎症环境中释放的17型型记忆细胞的非常大的储层，可能导致引发炎症性级联反应。通过调节HSCT之前的化学放射治疗引起的炎症可能会提供必要的启动信号，从而导致CD161HI CD8+ T细胞中TCR调节或分化的丧失，并允许它们为GVHD的发病机理做出贡献。该提案的具体目的是： 目的1。表征调节CD161HI TCM和TEM CD8+ T细胞中TCR信号转导的机制。这些研究将在健康个体的CD161HI CD8+ T细胞中定位TCR信号传导下调的位点，从而表明炎症性疾病中潜在的失调部位。 AIM 2。表征骨髓性同种异体HSCT后CD161HI CD8+ T细胞的恢复和功能，并确定该T细胞子集是否与GVHD有关。这些研究将确定骨髓性同种异体HSCT后CD161HI和CD161LO TCM和TEM CD8+细胞回收的动力学，如果血液或浸润组织中CD161HI CD8+细胞数量与急性GVHD相关。然后，我将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，以及来自HSCT患者CD161HI CD8+细胞中TCR信号的失调是否与GVHD相关。 职业发展和环境：指导的K99阶段将使我能够在我对CD161HI CD8+ T细胞的初步研究中进行的观察结果。具体而言，在K99期间，I将使用慢病毒载体进行基因重构和原代人T细胞中的敲低研究，以研究独特调控的CD161HI子集中的TCR信号传导所必需的技能。我将在Riddell博士和专家技术顾问Drs的指导下完成K99研究。 Tewari和Randolph-Habecker。获得K99阶段的数据和技能将有助于分析R00阶段中GVHD中CD161HI细胞的调节。 K99/R00阶段开发的技能将为未来的R01应用奠定基础，该应用集中在进一步了解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选择。免疫学计划由四名高级教职员工组成，他们在计划内外都有富有成效的合作历史。 FHCRC拥有出色的科学领导力，资源和出色的临床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，并确保我的K99/R00应用中提出的研究成功。 公共卫生相关性：申请人已经确定了一种新的方法来隔离和研究大部分免疫记忆细胞，这些免疫记忆细胞在疫苗接种或感染后提供保护。申请人在人类中发现的记忆T细胞对化学疗法具有抵抗力，这使他们可以在化学疗法治疗的患者中生存，并有助于从化学疗法中恢复后的免疫记忆重建为病毒。化学疗法 - 耐药细胞有可能引起严重的炎症，但由于严格的调节，在健康个体中不这样做。鉴定导致该调节损失的机制的研究可能会提供有关移植物与宿主疾病（GVHD）的原因和治疗的信息，血液干细胞移植的主要并发症以及其他炎症性或自身免疫性疾病。进一步的研究可能会使治疗的发展能够增加化学疗法或血干细胞移植后对感染和癌症的免疫力。