Endogenous glycosphingolipid antigens for NKT cells

NKT 细胞的内源性鞘糖脂抗原

• 批准号: 8277280
• 负责人: Dapeng Zhou
• 金额: $ 37.73万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-15 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8277280
• 关键词: Address; Adoptive Transfer; Agonist; Anabolism; Antigen-Antibody Complex; Antigen-Presenting Cells; Antigens; Area; Asthma; Autoimmune Diseases; Autoimmunity; Bacterial DNA; Biochemical; Biological Assay; Biology; CD1d antigen; Cell Line; Cells; Cellular Membrane; Ceramide glucosyltransferase; Characteristics; Clinical; Coenzymes; Communicable Diseases; Complex; Defect; Deficiency Diseases; Development; Disease; Enzymes; Event; Evolution; Fractionation; Generations; Genes; Genetic; Genomics; Glycolipids; Glycoside Hydrolases; Glycosphingolipids; Goals; Host Defense; Human; Human Genome; Human Genome Project; Hybrid Cells; Hybrids; Hypersensitivity; Immune; Immune System Diseases; Immune response; Immune system; Immunologic Receptors; Immunomodulators; Infection; Information Storage; Ions; Jaw; Knockout Mice; Knowledge; Leukocyte Adhesion Deficiency; Ligands; Link; Lipid Synthesis Pathway; Lipids; Lymphocyte; Malignant Neoplasms; Mass Fragmentography; Mass Spectrum Analysis; Memory; Metabolism; Molecular; Molecular Chaperones; Molecular Cloning; Mouse Strains; Mus; Natural Immunity; Natural Killer Cells; Outcome; Pathology; Pathway interactions; Pattern; Pharmaceutical Preparations; Physiological; Play; Proteins; Proteomics; RNA Interference; Receptor Signaling; Regulation; Research; Research Personnel; Research Training; Role; Series; Side; Source; Sphingolipid Activator Proteins; Staging; Stimulus; Structure; Surveys; System; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte and Natural Killer Cell; Technology; Testing; Thymus Gland; Tissues; Toll-like receptors; Tumor Antigens; UDP-galactose beta-D-galactosyl-1,4-glycosylceramide alpha-1,3-galactosyltransferase; Vertebrates; adaptive immunity; base; cell killing; cell type; chemical synthesis; embryonic antigen; experience; fight against; insight; isoglobotriaosylceramide; killer T cell; lipid metabolism; macrophage; mast cell; microbial; neutrophil; new technology; novel strategies; novel therapeutic intervention; novel therapeutics; pathogen; prevent; progenitor; receptor; success; vaccine development; viral DNA; viral RNA

Natural Killer T cells (NKT) are a hybrid cell type of NK cells and T cells, but are activated only by antigenic stimulation. NKT cells bridge innate and adaptive immunity through recognition of lipid ligands from both microbial and self origins. Endogenous NKT ligands, which are metabolism products of self lipids, are involved in immuno-pathology of infectious diseases, auto-immune diseases and cancer. However, molecular identification of these endogenous ligands has been difficult due to technical barriers. Recently, we found that NB-DGJ, a drug which inhibits glucosylceramide synthase, completely abolishes the development of NKT cells. More importantly, we identified isoglobotrihexosylceramide, one of the target glycosphingolipids being inhibited by NB-DGJ, as the first known natural ligand for NKT cells. However, iGb3 synthase knockout mice showed no defect in NKT cell development and function, indicating that other GSL and/or non-GSL ligands exist. Continued development of glycolipidomics assays, assisted by ion trap mass spectrometry technology, enhances our ability to further understand these mechanisms and identify new ligands. In this project, we will test the hypothesis that at least one endogenous ligand of NKT cells, other than iGb3, exists and is responsible for the development and activation of NKT cells. Identifying one or more endogenous ligands may permit new approaches to regulating the function of NKT cells in disease settings such as cancer and autoimmune diseases. The specific aims of this project are: 1) Determine the identities of stimulatory GSL and/or non-GSL antigens by biochemical fractionation and analysis; 2) Utilize the genetic pathways to identify potential endogenous NKT ligands. The molecular identification of natural ligands for NKT cells is not only an urgent, but also a challenging task in the field of innate immunity. Although the search for these ligands has lasted more than a decade, they remain elusive. We therefore have chosen a combined approach of biochemical fractionation/characterization and genetic knockdown of key enzymes to help bring this search to fruition. The completion of human genome project and the advancement of mass spectrometry technology provide the framework to move the field of lipid antigens forward and potentially permit the identification of new therapeutic approaches to some of the most complex immune-related diseases. More important, this project will provide early insights into cellular lipidomics studies, with a specific focus on the important immunological functions of glycolipids. More than 10% of the genes in the human genome are used for lipid metabolism; thus, knowledge and experience gained in this project may have profound implications on studies of functional lipidomics, an important but little-understood area compared with proteomics and genomics.

天然杀手T细胞（NKT）是NK细胞和T细胞的杂化细胞类型，但仅被激活 抗原刺激。 NKT细胞通过识别脂质配体来弥合先天和适应性免疫 来自微生物和自我起源。内源性NKT配体，它们是自我的代谢产物 脂质参与传染病，自身免疫性疾病和癌症的免疫病理学。 但是，由于技术，这些内源配体的分子鉴定很困难 障碍。最近，我们发现NB-DGJ是一种抑制葡萄糖酰胺合酶的药物，完全 废除NKT细胞的发展。更重要的是，我们鉴定 NB-DGJ抑制了靶糖磷脂的一种，这是第一个已知的天然配体 NKT细胞。然而，IGB3合酶基因敲除小鼠在NKT细胞发育中没有缺陷 功能，表明存在其他GSL和/或非GSL配体。继续发展 在离子陷阱质谱技术的协助下，糖脂素学测定法，增强了我们的能力 进一步了解这些机制并确定新的配体。 在这个项目中，我们将检验以下假设：NKT细胞的至少一个内源性配体 IGB3，存在并负责NKT细胞的发展和激活。识别一个或多个 内源配体可能允许调节NKT细胞功能的新方法 癌症和自身免疫性疾病等环境。该项目的具体目的是：1）确定 通过生化分级和分析的刺激GSL和/或非GSL抗原的身份； 2） 利用遗传途径鉴定潜在的内源性NKT配体。 NKT细胞的天然配体的分子鉴定不仅是紧急的，而且是一个具有挑战性的 先天免疫领域的任务。尽管对这些配体的搜索持续了 十年，他们仍然难以捉摸。因此，我们选择了生化的组合方法 分馏/表征和关键酶的遗传敲低以帮助带来此搜索 实现。人类基因组项目的完成和质谱的进步 技术提供了向前移动脂质抗原领域的框架，并有可能允许 鉴定一些与免疫相关疾病的新治疗方法。 更重要的是，该项目将提供对细胞脂肪态学研究的早期见解，并具有特定的 专注于糖脂的重要免疫功能。超过10％的基因 人基因组用于脂质代谢；因此，在这个项目中获得的知识和经验 可能对功能性脂肪态学的研究具有深远的影响，这是一个重要但鲜为人知的 与蛋白质组学和基因组学相比。