HLA-D ANTIGENS IN INSULIN-DEPENDENT DIABETES

胰岛素依赖型糖尿病中的 HLA-D 抗原

• 批准号: 3447572
• 负责人: DIMITRI S MONOS
• 金额: $ 5.77万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3447572
• 关键词: B lymphocyte; antibody specificity; gene expression; genetic manipulation; high performance liquid chromatography; histocompatibility antigens; human subject; immunogenetics

The objectives of this study are 1) to biochemically identify the specific gene products associated with the HLA-D region that contribute to the susceptibility of Insulin Dependent Diabetes (IDD) and 2) to study the expression of the biochemically identified forms, as IDD specific on transfectant cells. This will be accomplished by using cryopreserved cells and sera from a large number of patients that have been collected and typed for HLA class I and II antigens. In addition, a panel of 23B-lymphoid cell lines (B-LCL) derived from 10 families will be utilized. The study will consist of: 1) Characterization of HLA-DR and DO alpha and beta subunits from B-LCL derived from IDD patient carrying the HLA-DR3 and/or DR4 haplotypes by two dimensional polyacrylamide gel electrophoresis (2D-PAGE), high pressure liquid chromatography (HPLC) and restriction fragment analysis of the DNA (RFLP). 2) Identification of combinatorial (hybrid) DQ antigens forming unique Ia determinants generated as a result of the association of the alpha and beta chains from two haplotypes which can function as restriction elements conferring susceptibility to IDD. 3) Identification of alloantibodies reactive with IDD associated antigens in the sera of multiparous mothers of IDD patients. 4) Isolation of the genes coding for the molecular forms of DR or DO that the biochemical studies indicated to be IDD specific and generation of transfectants expressing these genes. Information generated from this study will help to identify molecular forms more frequently seen among IDD patients and indirectly to use them as markers for identifying those at risk. The transfectants generated will provide an excellent system for studying the genetics of the susceptibility to IDD and will also constitute a model for other Ia associated diseases.

这项研究的目标是：1)生化鉴定 与人类白细胞抗原-D区相关的特定基因产物 胰岛素依赖型糖尿病的易感性 (IDD)和2)表达的生化研究 已鉴定的形式，如转基因细胞上特有的IDD。这将是 通过使用冷冻保存的细胞和来自 收集了大量患者并对其进行了分类 人类白细胞抗原I、II类。此外，一组23B-淋巴 将利用来自10个家系的细胞系(B-LCL)。这个 研究内容包括：1)人类白细胞抗原DR和DO的特性 IDD患者B-LCL的α和β亚基 二维携带人类白细胞抗原-DR3和/或DR4单倍型 高压聚丙烯酰胺凝胶电泳法(2D-PAGE) 高效液相色谱和限制性内切酶片段分析 DNA(RFLP)。2)组合(混杂)的识别 由此产生的形成唯一Ia决定簇的DQ抗原 α链和β链的相互作用 可作为限制因素的单倍型 对IDD的易感性。3)同种异体抗体反应性鉴定 经产妇血清中存在IDD相关抗原 在碘缺乏病患者中。4)编码基因的分离 DR或DO的分子形式的生化研究 被指示为IDD特异性和转染体的产生 表达这些基因。 这项研究产生的信息将有助于确定 分子形式更常见于IDD患者和 间接地将它们用作识别风险人群的标记。 产生的转染体将提供一个很好的系统 研究IDD易感性的遗传学，还将 建立了其他Ia相关疾病的模型。