MHC CLASS II STRUCTURE STUDIES

MHC II 类结构研究

• 批准号: 6201295
• 负责人: DON C WILEY
• 金额: $ 17.41万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6201295
• 关键词: MHC class II antigen; NOD mouse; SDS polyacrylamide gel electrophoresis; T cell receptor; X ray crystallography; autoantigens; chemical structure; diabetes mellitus genetics; disease /disorder proneness /risk; gene expression; genetically modified animals; high performance liquid chromatography; insulin dependent diabetes mellitus; molecular cloning; polymerase chain reaction; radionuclides; receptor binding

Specific MHC class II molecules are linked to the susceptibility to autoimmune diseases in humans and experimental animals. The MHC class II molecules HLA-DQ8 and HLA-DQ2 in humans and I-Ag7 in NOD mice, for example, appear to confer susceptibility to type I diabetes. The specificity of these molecules is found in their peptide binding sites. The three-dimensional structures of MHC class II complexes with peptides from candidate auto antigens could provide a description of the atomic interactions specific to the high-risk class II molecules. A detailed understanding of the specificity of these MHC molecules should provide a background for helping to discover antigenic epitopes from candidate autoantigens and guide the design of selective blockers of these MHC class II molecules. The synthesis of specific blockers would allow the mechanisms by which these MHC molecules cause susceptibility to be examined in the initiation and progression of disease. The specific aims of this proposal are: 1) to express soluble, human MHC class II molecules (HLA-DQ8, HLA-DQ2) conferring susceptibility to diabetes and to load them with peptides from candidate autoantigens, 2) to crystallize HLA-DQ8 and HLA-DQ2 complexed with peptides and determine their three-dimensional structures by X-ray crystallography, 3) to use structural information to design both specific ligands and libraries of non-peptidic ligands of the peptide binding site of MHC class II molecules conferring susceptibility to diabetes, 4) to assay these compounds for their ability to bind to their class II receptor, and 5) to examine the effect of selective blockers on the development of autoaggressive T cells and on the progression of disease in NOD mice transgenic for DQ8 or DQ2 susceptibility genes.

特定 MHC II 类分子与对以下疾病的易感性相关： 人类和实验动物的自身免疫性疾病。 MHC II 类 人类的 HLA-DQ8 和 HLA-DQ2 分子以及 NOD 小鼠的 I-Ag7 分子， 例如，似乎会导致对 I 型糖尿病的易感性。这 这些分子的特异性在于它们的肽结合位点。 MHC II类肽复合物的三维结构 候选自身​​抗原可以提供原子的描述 高风险 II 类分子特有的相互作用。详细的 了解这些 MHC 分子的特异性应该提供一个 帮助从候选者中发现抗原表位的背景 自身抗原并指导这些 MHC 类别的选择性阻断剂的设计 II 分子。特定阻断剂的合成将允许 这些 MHC 分子导致易感性的机制 检查疾病的发生和进展。 该提案的具体目标是：1）表达可溶性人类 MHC II 类分子（HLA-DQ8、HLA-DQ2）赋予对 糖尿病并用候选自身抗原的肽加载它们，2) 结晶与肽复合的 HLA-DQ8 和 HLA-DQ2 并测定 通过 X 射线晶体学分析它们的三维结构，3) 使用 用于设计特定配体和库的结构信息 MHC II 类分子肽结合位点的非肽配体 赋予糖尿病易感性，4) 测定这些化合物 它们与 II 类受体结合的能力，以及 5) 检查 选择性阻断剂对自身攻击性 T 细胞发育的影响 以及 DQ8 或 DQ2 转基因 NOD 小鼠疾病进展的影响 易感基因。