Self-glycolipid-reactive T cells: repertoire & function

自身糖脂反应性 T 细胞：全部

• 批准号: 6828467
• 负责人: Vipin Kumar
• 金额: $ 35.92万
• 依托单位: TORREY PINES INST FOR MOLECULAR STUDIES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6828467
• 关键词: CD1 molecule; T lymphocyte; autoimmunity; cellular immunity; cerebrosides; cytokine; drug screening /evaluation; enzyme linked immunosorbent assay; genetically modified animals; glycolipids; hybridomas; immune response; laboratory mouse; leukocyte activation /transformation; myelinopathy; natural killer cells; nervous system disorder chemotherapy; neutralizing antibody; nucleic acid sequence; passive immunization; phenotype; polymers; tissue /cell culture

DESCRIPTION (provided by applicant): Some T cells recognize lipid antigens presented by class I MHC-like CD1 molecules. Although self-lipid antigens activating them have not yet been identified, most NK T cells are thought to be autoreactive and can be rapidly activated by a surrogate glycolipid ligand alpha-galactosyl ceramide or alpha-GalCer. To characterize self-glycolipid-reactive T cells, their phenotype and their physiological function, we have focused on the recognition of a major myelin-derived glycolipid, sulfatide, (3'-sulfated beta-galactosyl ceramide). We have generated sulfatide-CD1d-tetramers and have identified sulfatide-reactive T cell populations in the liver, thymus and spleen from naive C57BL/6 mice. We have also generated sulfatide-reactive T cell hybridomas. During the course of antigen-induced experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis (MS) in humans, sulfatide-CD1d-tetramer+ T cells are selectively increased several-fold within the central nervous system. Treatment with sulfatide can both prevent and reverse ongoing EAE in wild type but not in CD1d-deficient mice. Based on our findings we propose that sulfatide is a self-glycolipid ligand recognized by a distinct population of CD1d-restricted T cells that can be activated to modulate autoimmune responses. We will use sulfatide- and a-GalCer-CD1d-tetramers and antibodies against various cell surface markers on NK T cells to characterize the phenotype of sulfatide-reactive T cells. Sulfatide-reactive T cell hybridomas will be used to analyze the antigen fine specificity and the TCR V-gene repertoire. Using glycolipid tetramers and Elispot analysis we will determine the dynamics of their activation and investigate mutual interactions among different lipid-reactive T cell populations. We will determine the number, phenotype and the fate of NK cells, B cells and myelin protein-reactive pathogenic CD4 T cells following sulfatide injection of wild type and CD1d-deficient mice. In adoptive transfer experiments we will directly examine the regulatory potential of sulfatide-CD ld-tetramer+ T cells. Neutralizing antibodies and mice genetically deficient for type 1 and 2 cytokines will be used to determine their role in the modulation of EAE by sulfatide. We will optimize treatment of ongoing disease with sulfatide in order to explore its therapeutic potential. These studies are important not only for understanding the biology of a naturally occurring self-glycolipid-reactive T cell population, but also because of the highly conserved nature of CD1d molecules across species they will form the basis for manipulation of human autoimmune demyelinating diseases, such as MS.

描述（由申请人提供）：一些T细胞识别由I类MHC样CD1分子提出的脂质抗原。尽管尚未鉴定出激活它们的自脂抗原，但大多数NK T细胞被认为具有自动反应性，并且可以通过替代糖糖脂质配体α-甲乳糖基神经酰胺或α-α迅速激活。为了表征自糖反应性T细胞的表型和生理功能，我们专注于识别主要的髓磷脂衍生的糖脂硫化物（3'-硫化的β-半乳糖基酰胺）。我们已经产生了硫化物-CD1D四聚体，并从天真的C57BL/6小鼠中鉴定出肝脏，胸腺和脾脏中的磺胺反应性T细胞群体。我们还产生了磺胺反应性T细胞杂交瘤。在抗原诱导的实验性自身免疫性脑脊髓炎（EAE）过程中，一种在人类中多发性硬化症（MS）的模型，硫化物-CD1D-四聚体+ T细胞在中枢神经系统中选择性增加了几倍。用亚磺胺治疗可以预防和反向野生型的EAE，但在CD1D缺乏小鼠中不能预防和反向。根据我们的发现，我们建议硫化物是一种由不同的CD1D限制T细胞群识别的自糖脂配体，可以激活以调节自身免疫反应。我们将使用硫化物和A-甘油-CD1D四聚体以及针对NK T细胞上各种细胞表面标记的抗体来表征磺胺反应性T细胞的表型。硫化物反应性T细胞杂交瘤将用于分析抗原的精细特异性和TCR V-Gene曲目。使用糖脂四聚体和ELISPOT分析，我们将确定其激活的动力学，并研究不同脂质反应性T细胞种群之间的相互作用。我们将在硫化物注射野生型和CD1D缺陷小鼠后确定NK细胞，B细胞和髓磷脂蛋白反应性致病CD4 T细胞的数量，表型和命运。在收养转移实验中，我们将直接检查硫化物-CD LD-四聚体+ T细胞的调节潜力。中和抗体和小鼠在遗传上缺乏1型和2个细胞因子的缺乏来确定它们在硫化物调节EAE中的作用。我们将用亚硫酸盐优化正在进行的疾病的治疗，以探索其治疗潜力。这些研究不仅对于理解天然存在的自糖脂质反应性T细胞群的生物学至关重要，而且还因为跨物种的CD1D分子的高度保守性质，它们将成为操纵人类自身免疫性脱氧疾病的基础，例如MS。