HLA-D ANTIGENS IN INSULIN-DEPENDENT DIABETES

本研究的目的是：1）生物化学鉴定 与HLA-D区域相关的特定基因产物， 导致胰岛素依赖型糖尿病的易感性 (IDD)和2）研究表达的生化 鉴定的形式，如IDD特异性转染细胞。 这将 通过使用冷冻保存的细胞和血清， 大量的病人已经被收集和分类， HLA I类和II类抗原。 此外，一组23 B淋巴样 将使用来自10个家族的细胞系（B-LCL）。 的 研究将包括：1）HLA-DR和DO的表征 来自IDD患者的B-LCL的α和β亚单位 携带HLA-DR 3和/或DR 4单倍型， 聚丙烯酰胺凝胶电泳（2D-PAGE），高压 液相色谱（HPLC）和限制性片段分析 DNA（RFLP） 2)组合（杂交）的鉴定 DQ抗原形成作为结果产生的独特Ia决定簇 从两个不同的基因中 单倍型可以作为限制性元件， 易患碘缺乏病。 3)同种抗体反应性鉴定 经产母亲血清中IDD相关抗原 IDD患者。 4)分离编码以下的基因： DR或DO的分子形式， 表明是IDD特异性的，并产生转染子 表达这些基因。 本研究产生的信息将有助于确定 分子形式在IDD患者中更常见， 间接地将其用作识别风险人群的标记。 所产生的转染子将提供一个极好的系统， 研究IDD易感性的遗传学，并将 构成了其他Ia相关疾病的模型。

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.