Self-glycolipid-reactive T cells: repertoire & function

自身糖脂反应性 T 细胞：全部

• 批准号: 7226324
• 负责人: Vipin Kumar
• 金额: $ 35.38万
• 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 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7226324
• 关键词: Adoptive Transfer; Amino Acids; Antibodies; Antigens; Autoimmune Process; Autoimmune Responses; B-Lymphocytes; Binding; Biology; C57BL/6 Mouse; CD1 Antigens; CD3 Antigens; CD4 Positive T Lymphocytes; CD44 gene; CD8B1 gene; Cell surface; Cells; Ceramides; Characteristics; Complex; Demyelinating Diseases; Demyelinations; Disease; Encephalomyelitis; Enzyme-Linked Immunosorbent Assay; Experimental Autoimmune Encephalomyelitis; Figs - dietary; Galactosylceramides; Genes; Glycolipids; Homologous Gene; Human; Hybridomas; Immunity; Infiltration; Injection of therapeutic agent; Inorganic Sulfates; Interferons; Interleukin-10; Interleukin-13; Interleukin-4; Label; Length; Ligands; Lipids; Liver; MHC Class I Genes; Mediating; Membrane; Memory; Modeling; Multiple Sclerosis; Mus; Myelin; Myelin Proteins; Natural Killer Cells; Nature; Neuraxis; Numbers; PTPRC gene; Peptides; Phenotype; Physiological; Population; Population Heterogeneity; Prevention; Proliferating; Reagent; Reverse Transcriptase Polymerase Chain Reaction; Role; Sorting - Cell Movement; Source; Specificity; Spleen; Splenocyte; Staining method; Stains; Standards of Weights and Measures; Sulfoglycosphingolipids; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Therapeutic Uses; Thinking; Thymus Gland; Transgenic Mice; Unspecified or Sulfate Ion Sulfates; alpha-galactosylceramide; base; cytokine; enzyme linked immunospot assay; in vivo; killer T cell; neutralizing antibody; prevent; research study; response

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分子呈递的脂质抗原。尽管尚未确定激活它们的自身脂质抗原，但大多数 NK T 细胞被认为具有自身反应性，并且可以被替代糖脂配体 α-半乳糖基神经酰胺或 α-GalCer 快速激活。为了表征自身糖脂反应性 T 细胞、其表型和生理功能，我们重点关注了主要髓磷脂衍生糖脂、硫苷脂（3'-硫酸化 β-半乳糖基神经酰胺）的识别。我们已经生成了硫苷脂-CD1d-四聚体，并鉴定了幼稚 C57BL/6 小鼠的肝脏、胸腺和脾脏中的硫苷脂反应性 T 细胞群。我们还生成了硫苷脂反应性 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）（一种人类多发性硬化症（MS）模型）的过程中，硫苷脂-CD1d-四聚体+ T细胞在中枢神经系统内选择性地增加数倍。硫苷脂治疗可以预防和逆转野生型小鼠中正在进行的 EAE，但不能用于 CD1d 缺陷小鼠。根据我们的研究结果，我们提出硫苷脂是一种自身糖脂配体，可被特定的 CD1d 限制性 T 细胞群识别，可被激活以调节自身免疫反应。我们将使用硫苷脂-和 a-GalCer-CD1d-四聚体以及针对 NK T 细胞上各种细胞表面标记物的抗体来表征硫苷脂反应性 T 细胞的表型。硫脂反应性 T 细胞杂交瘤将用于分析抗原精细特异性和 TCR V 基因库。使用糖脂四聚体和 Elispot 分析，我们将确定其激活的动态并研究不同脂质反应性 T 细胞群之间的相互作用。我们将确定野生型和 CD1d 缺陷小鼠注射硫苷脂后 NK 细胞、B 细胞和髓磷脂蛋白反应性致病性 CD4 T 细胞的数量、表型和命运。在过继转移实验中，我们将直接检查硫苷脂-CD 1d-四聚体+ T 细胞的调节潜力。中和抗体和遗传缺陷 1 型和 2 型细胞因子的小鼠将用于确定它们在脑硫苷脂调节 EAE 中的作用。我们将优化硫脑苷脂对正在进行的疾病的治疗，以探索其治疗潜力。这些研究不仅对于了解天然存在的自身糖脂反应性 T 细胞群的生物学非常重要，而且由于 CD1d 分子在不同物种之间的高度保守性，它们将构成治疗人类自身免疫性脱髓鞘疾病（如多发性硬化症）的基础。