Study on the Pathogenesis of Type 1 Diabetes Mellitus

1型糖尿病发病机制研究

基本信息

批准号：
01480290
负责人：
HANAFUSA Toshiaki
金额：
$ 4.29万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480290/
关键词：
Type 1 diabetes MHC antigens Cytokines Interferon-gamma Pancreatic beta cells Pathogenesis Insulin secretion Molecular biology インスリノ-マ細胞トランスフェクション遺伝子導入 FACS 腫瘍壊死因子 Northern blotting

项目摘要

The pathogenic role of abnormal expression of major histocompatibility complex (MHC) antigens on pancreatic beta cells was studied using molecular biological techniques.We first tested which cytokines could induce MHC antigen expression on rat insulinoma cell line RINm5F. Messenger RNAs of class I and class II MHC molecules were examined by Northern analysis. The result showed that, in the absence of cytokines, 1) class I MHC antigen mRNA was constitutively expressed but 2) class II MHC mRNA was not detected in RINm5F cells. Class I mRNA was increased in response to either interferonーgamma (IFNgamma) or IFNgamma plus tumor necrosis factorーalpha (TNFalpha). However, no class II mRNA was detected after the challenge of any cytokines. On the other hand, insulin mRNA was decreased in the presence of either IFNgamma or IFNgamma+TNFalpha.We secondly examined the effect of MHC antigen expression on beta cell function, namely insulin secreting capacity. RINm5F cells were transfected with human MHC class I antigen (HLA-Cw2) gene and twelve clones with different degrees of expression of the transfectant were obtained. Insulin secreting capacity of the clones in response to glucose was negatively correlated with the amount of class I MHC antigen expressed.These results indicate that the expression of MHC antigen in beta cells is controlled by various cytokines and that insulin secreting capacity is impaired by the expression of MHC antigens in beta cells. This suggests the importance of MHC antigen expression in the pathogenesis of Type 1 diabetes mellitus.

使用分子生物学技术研究了主要的组织相容性复合物（MHC）抗原在胰腺β细胞上异常表达的致病作用。我们首先测试哪种细胞因子可以在大鼠胰岛素瘤细胞系RINM5F上诱导MHC抗原表达。通过北方分析检查了I类和II类MHC分子的Messenger RNA。结果表明，在没有细胞因子的情况下，1）I类MHC抗原mRNA始终表达，但2）在RINM5F细胞中未检测到II类MHC mRNA。 I类mRNA因对干扰素 - 伽马（IFNGAMMA）或IFNGAMMA加肿瘤坏死因子-Alpha（TNFalpha）而增加。但是，在任何细胞因子的挑战之后，未检测到II类mRNA。另一方面，在存在IFNGAMMA或IFNGAMMA+TNFALPHA的情况下，胰岛素mRNA降低了。我们其次研究了MHC抗原表达对β细胞功能的影响，即胰岛素分泌能力。用人MHC I类抗原（HLA-CW2）基因翻译RINM5F细胞，并获得了具有不同程度的翻译表达程度的克隆。克隆对葡萄糖的胰岛素分泌能力与I类MHC抗原表达的量负相关。这些结果表明，MHC抗原在β细胞中的表达受到各种细胞因子的控制，并且胰岛素分泌能力受到β细胞中MHC抗原的表达损害。这表明MHC抗原表达在1型糖尿病的发病机理中的重要性。