Immune Recognition of Glycolipids

糖脂的免疫识别

• 批准号: 7329680
• 负责人: Luc Teyton
• 金额: $ 53.11万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7329680
• 关键词: Address; Agonist; Anti-Bacterial Agents; Antibacterial Response; Antigen Targeting; Antigen-Presenting Cells; Antigens; Autoimmunity; Binding; Biochemical; Biochemistry; Biological; CD1 Antigens; Carrier Proteins; Cells; Cellular Immunology; Cellular biology; Ceramides; Chemicals; Chemistry; Classification; Collaborations; Complement; Complex; Effector Cell; Elements; Evolution; Fluorescence Resonance Energy Transfer; G(M2) Activator Protein; Galactosylceramides; Genetic Polymorphism; Glycolipids; Goals; Head; Human; Immune; Immune system; In Vitro; Inflammation; Knockout Mice; Knowledge; Ligand Binding; Ligands; Lipid Binding; Lipids; Lipoproteins; Lysosomes; Measurement; Modification; Mus; Mutagenesis; Nature; Numbers; Pathway interactions; Peptides; Physiology; Population; Principal Investigator; Protein Engineering; Proteins; Publications; Reaction; Recombinant Proteins; Role; SAP-A Protein; Shapes; Solutions; Structure; Surface; System; T-Cell Receptor; T-Lymphocyte; Thermodynamics; Thymus Gland; Time; Ursidae Family; Vaccine Antigen; Work; base; design; experience; extracellular; immunogenic; in vivo; isoglobotriaosylceramide; lipid transfer protein; lipid transport; oxidation; particle; programs; protein expression; receptor; reconstitution; research study; response; skills; success; trafficking; uptake

The project entitled "Immune recognition of glycolipids" fits into a collaborative Program Project by addressing biochemical and structural questions that complement biological studies led by Dr.Bendelac, and by using extensively chemical compounds developed in collaboration with Dr.Savage. Our 3 Specific Aims will be: Lipid Transfer Proteins (LTPs). We will examine the contribution of saposins A through D, Niemann-Pick type C-2 and GM2 activator protein in the shaping of the repertoire of lipids presented by CD1 molecules in professional antigen presenting cells. In vivo studies in knock-out mice will evaluate the contribution of each protein in the selection of NKT cells as well as in anti-bacterial responses. In vitro studies will attempt to reconstitute the complex situation of the lysosome where CD1 resides with a mixture of small LTPs and a wide spectrum of glycolipids. Biophysical measurements and structural studies will also attempt to understand the mechanisms of transfer of lipids between CD1 and LTPs using real time FRET and SPR. Extracellular Lipid Transport and Uptake. Extracellular lipids are kept in solution by associating with lipoproteins particles or lipid transport proteins. The delivery of immunogenic lipids, such as bacterial lipids, follows some of the same trafficking pathways. We will determine which pathway are important for known agonists of NKT cells such as a-galactosyl ceramide (a-GalCer) and bacterial glucuronyl ceramides. The identification of transport proteins will be driven by the usage of biotinylated compounds and biochemistry. Lipid uptake and trafficking to the lysosome will be determined by cell biology and the usage of knock-out mice for surface receptors. The consequences of in vivo modifications, such as oxidation that accompanies inflammation, will also be studied with respect to transport. Finally, we will attempt some of the knowledge gained by this specific aim to devise new strategies for delivering lipid antigens in vivo. Structural Studies of CD1 recognition by Va14 T cell receptors. We know from experience that this approach is highly unpredictable but we also know that it is unlikely that a sustained effort will not deliver some results. This approach has allowed us to determine important structures such as CD1 bound to a- GalCer but more structures are required. The main goal of this aim is to understand the ability of Va14 TCRs to recognize ligands as different as CD1-iGb3 and CD1.-a-GalCer. To attain this goal new protein engineering is required and will be exposed in details. Strong preliminary results will help us in the design of these experiments. Additional CD1-lipid structures studies will also be proposed to understand the influence of acyl chain unsaturation and head group modifications. Finally, we will also carry the systematic mutagenesis of the Va14 chain to understand the thermodynamics of the reaction Va14/CD1.

该项目标题为“免疫识别糖脂”，适合于一个协作计划项目。 解决由Bendelac博士领导的生物学研究的生化和结构性问题，以及 通过使用与Ssavage博士合作开发的广泛化合物。我们的三个特定目标 将是：脂质转移蛋白（LTPS）。我们将研究saposins a至d的贡献， Niemann-Pick型C-2和GM2激活蛋白在CD1提出的脂质曲目的构造中 专业抗原呈递细胞中的分子。淘汰小鼠中的体内研究将评估 每种蛋白在选择NKT细胞以及抗细菌反应中的贡献。体外 研究将试图重建CD1与混合物驻留的溶酶体的复杂情况 小的LTP和广泛的糖脂。生物物理测量和结构研究也将 尝试使用实时fret了解CD1和LTP之间脂质转移的机制 和spr。细胞外脂质转运和摄取。通过关联，将细胞外脂质保存在溶液中 含脂蛋白颗粒或脂质转运蛋白。免疫原性脂质的递送，例如细菌 脂质遵循一些相同的贩运途径。我们将确定哪种途径对于 NKT细胞的已知激动剂，例如A-半乳糖苷神经酰胺（A-Galcer）和细菌葡萄糖酰胺。 运输蛋白的鉴定将由生物素化合物的使用驱动 生物化学。脂质的摄取和对溶酶体的贩运将由细胞生物学确定和使用 表面受体的敲除小鼠。体内修饰的后果，例如氧化 伴随炎症，还将研究运输。最后，我们将尝试一些 这种特定目的获得的知识旨在制定在体内提供脂质抗原的新策略。 VA14 T细胞受体识别CD1的结构研究。我们从经验中知道这一点 方法是不可预测的，但我们也知道，持续的努力不可能无法实现 一些结果。这种方法使我们能够确定重要的结构，例如与A-结合的CD1 galcer，但需要更多的结构。此目标的主要目标是了解VA14 TCR的能力 识别与CD1-IGB3和CD1.-A-Galcer不同的配体。要达到这个目标新蛋白 需要工程，并将详细介绍。强大的初步结果将有助于我们设计 这些实验。还将提出其他CD1-脂质结构研究以了解影响 酰基链的不饱和度和头部组修饰。最后，我们还将进行系统的 VA14链的诱变，以了解反应VA14/CD1的热力学。