Immune Recognition of Glycolipids

糖脂的免疫识别

基本信息

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
项目类别：
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

项目摘要

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.

题为“糖脂的免疫识别”的项目属于以下合作项目：解决生化和结构问题，补充 Bendelac 博士领导的生物学研究，以及通过广泛使用与 Savage 博士合作开发的化合物。我们的 3 个具体目标将是：脂质转移蛋白（LTP）。我们将检查皂苷 A 至 D 的贡献， Niemann-Pick C-2 型和 GM2 激活蛋白在 CD1 呈递的脂质库的塑造中专职抗原呈递细胞中的分子。基因敲除小鼠的体内研究将评估每种蛋白质在 NKT 细胞选择以及抗菌反应中的贡献。体外研究将尝试重建 CD1 与混合物所在的溶酶体的复杂情况小 LTP 和广谱糖脂。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。细胞外脂质运输和摄取。细胞外脂质通过缔合而保持在溶液中与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。我们将确定哪条途径对于已知的 NKT 细胞激动剂，例如α-半乳糖神经酰胺（α-GalCer）和细菌葡萄糖醛酸神经酰胺。转运蛋白的鉴定将通过生物素化化合物的使用来驱动生物化学。脂质的摄取和运输到溶酶体将由细胞生物学和使用来决定表面受体基因敲除小鼠。体内修饰的后果，例如氧化伴随炎症，也将在运输方面进行研究。最后，我们将尝试一些通过这一特定目标获得的知识旨在设计在体内递送脂质抗原的新策略。 Va14 T 细胞受体 CD1 识别的结构研究。我们从经验中知道，这方法是高度不可预测的，但我们也知道，持续的努力不太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，但需要更多结构。该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。为了实现这一目标新蛋白质工程是必需的，并将详细公开。强有力的初步结果将帮助我们设计这些实验。还将提出额外的 CD1-脂质结构研究，以了解其影响酰基链不饱和度和头基修饰。最后，我们还将进行系统的 Va14 链的诱变，以了解 Va14/CD1 反应的热力学。