A novel population of PLZF+CD8+ regulatory T cells: phenotype and function

PLZF CD8 调节性 T 细胞的新群体：表型和功能

• 批准号: 10247312
• 负责人: Vipin Kumar
• 金额: $ 55.77万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-17 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247312
• 关键词: Adoptive Transfer; Apoptosis; Area; Arthritis; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Autoimmunity; Biology; Blood; Blood Cells; Bone Marrow; CD 200; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cells; Characteristics; Clinical Research; Colitis; Comparative Study; Cross Presentation; Data; Development; Disease; Disease model; Experimental Autoimmune Encephalomyelitis; FOXP3 gene; Feedback; Foundations; Frequencies; Future; Gene Expression Profile; Genetic; Genetic Transcription; Goals; Grant; Granzyme; Homeostasis; Human; IL2RB gene; ITGAX gene; Immune; Immune Tolerance; Immunization; Immunotherapy; Individual; Inflammation; Inflammatory; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Interleukin-15; Investigation; KLRB1 gene; Liver; Lupus; Lymphocyte; Lymphoid Tissue; Maintenance; Mediating; Memory; Microglia; Modeling; Molecular; Mus; Outcome; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Physiological; Play; Population; Qa-1 Antigen; Rag1 Mouse; Recovery; Regulation; Regulatory T-Lymphocyte; Research; Resistance; Rheumatoid Arthritis; Role; Signal Transduction; Signaling Molecule; Surface; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Time; Tissues; Work; ZNF145 gene; autoreactivity; base; cytokine; cytotoxic CD8 T cells; design; in vitro Assay; molecular marker; mouse model; novel; novel therapeutics; overexpression; perforin; peripheral blood; programs; single-cell RNA sequencing; tissue injury; transcription factor; transcriptome

Immune tolerance mediated by regulatory T cells (Treg) is important in the control of inflammatory and autoimmune diseases. The detailed biology of CD8 Treg is lacking due to the absence of molecular markers to distinguish them from conventional CD8+ T cells (CD8conv), as Foxp3 expression is for CD4 Treg. We have discovered that the expression of the promyelocytic leukemia zinc finger (PLZF) transcription factor together with other markers distinguishes a novel, unconventional CD8 Treg population from other CD8conv as well as unconventional T cells. Though, CD8 Treg are enriched in liver of naïve mice, they are also present in other lymphoid tissues and in human PBMC and display an activated/memory phenotype and attributes of innate-like T cells. Our long-term goal is to characterize the biology of CD8 Treg in both mice and humans and to understand the negative feedback regulatory mechanism(s) employed by CD8 Treg that limits excessive immune stimulation. These studies are highly significant as they will have potential to not only uncover a key CD8 Treg-mediated molecular mechanism(s) involved in immune homeostasis, but also will have major implications for the development of new immune therapies in autoimmune diseases. The objective of this grant is to characterize the transcriptome signature, development, activation/expansion and mechanism of regulation of CD8 Treg in murine models and also to determine their molecular and suppressive functions in humans. The central hypothesis is that the PLZF transcription program allows acquisition of innate-like and memory features in CD8 Treg that enables them to control promptly and effectively inflammatory autoimmune responses by targeting activated T cells as well as APCs. Based upon our critical preliminary data that indicate a unique phenotype of CD8 Treg (PLZF+TCRαβ+CD8αα+), we propose the following specific aims: Aim 1. To determine their unique transcriptional program and molecular pathways, single cell RNA sequencing analysis of CD8 Treg subsets will be carried out in both the periphery and in the inflamed CNS. A bone marrow chimeric approach will be used to determine the role of PLZF transcription factor in their development and function; Aim 2. To determine the critical role of the cross-presentation mechanism in the physiological induction of CD8 Treg, mice genetically lacking cDC1 will be used. Genetic depletion of CD8 Treg in PLZFF/F mice will decipher their role in the recovery and resistance from experimental autoimmune encephalomyelitis. A critical role for the regulatory molecules, including CD200 and Fgl2, in the inhibition of DC function and/or Th1/Th17 induction will be elucidated; and Aim 3. To characterize the transcriptional signature and suppressive mechanism(s) involved in human CD8 Treg, sorted cells from peripheral blood of healthy individuals will be used. Collectively, our proposed studies will have a major impact in the emerging field of unconventional T cells and in the CD8 Treg-mediated regulatory mechanism(s) involved in the maintenance of immune tolerance.

由调节性T细胞（Treg）介导的免疫耐受在控制炎症和免疫耐受中是重要的。 自身免疫性疾病由于缺乏分子标记物，CD 8 Treg的详细生物学缺乏。 将它们与常规的CD 8 + T细胞（CD 8 conv）区分开，因为Foxp 3表达用于CD 4 Treg。我们有 发现早幼粒细胞白血病锌指（PLZF）转录因子的表达与 其他标志物将新的、非常规的CD 8 Treg群体与其他CD 8 conv区分开， 非常规T细胞虽然CD 8 Treg在未处理小鼠的肝脏中富集，但它们也存在于其他小鼠中。 淋巴组织和人PBMC中，并显示出激活/记忆表型和先天性类 T细胞。我们的长期目标是描述小鼠和人类中CD 8 Treg的生物学特征，并了解其在小鼠和人类中的作用。 由CD 8 Treg采用的限制过度免疫刺激的负反馈调节机制。 这些研究非常重要，因为它们不仅有可能揭示关键的CD 8 Treg介导的细胞凋亡， 参与免疫稳态的分子机制，但也将对免疫系统产生重大影响。 自身免疫性疾病的新免疫疗法的发展。该补助金的目的是描述 CD 8调节性T细胞的转录组特征、发育、活化/扩增及其调控机制 小鼠模型，并确定其在人类中的分子和抑制功能。中央 一种假设是PLZF转录程序允许在CD 8中获得先天样和记忆特征， Treg，使他们能够迅速有效地控制炎症性自身免疫反应， 活化的T细胞和APC。根据我们的关键初步数据，表明一种独特的表型， CD 8调节性T细胞（PLZF+TCRαβ+ CD 8 αα+），我们提出以下具体目标：目的1.以确定它们独特的 转录程序和分子途径，CD 8 Treg亚群的单细胞RNA测序分析将 在外周和发炎的中枢神经系统中进行。骨髓嵌合体方法将用于 明确PLZF转录因子在其发育和功能中的作用;目的2.确定临界 交叉呈递机制在CD 8调节性T细胞生理诱导中的作用， 将使用cDC 1。PLZFF/F小鼠中CD 8调节性T细胞的遗传耗竭将解释其在恢复中的作用， 实验性自身免疫性脑脊髓炎对于调节分子来说， 包括CD 200和Fg 12在抑制DC功能和/或Th 1/Th 17诱导中的作用将被阐明; 3.为了表征人CD 8 Treg中涉及的转录特征和抑制机制， 将使用来自健康个体外周血的分选细胞。总的来说，我们提出的研究将有 在新兴的非常规T细胞领域和CD 8 Treg介导的调节 参与维持免疫耐受的机制。