Analysis of islet antoantigens by the screening of a mouse islet cDNA expression library.

通过筛选小鼠胰岛 cDNA 表达文库来分析胰岛抗原。

• 批准号: 07671166
• 负责人: YAMADA Kentaro
• 金额: $ 1.6万
• 依托单位: Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671166/
• 关键词: Type 1 diabetes; NOD mouse; islet autoantigen; chromogranin A; I型糖尿病

Two clones were obtained by immunoscreening of an NOD mouse islet lambdagt11 expression library with prediabetic NOD mouse sera. The clones (C17 and C18) were recognized by NOD mouse sera but not by Balb/c mouse sera. Antibodies against C17 and C18 fusion proteins were detected in 80% and 10% of sera from 10 week-old female NOD mice, respectively. DNA sequences of C18 were identical to mouse chromogranin A cDNA (596-1892). Nucleotide sequences of C17 showed a poly (A) tail and open reading frame for 46 amino acids. C17 mRNA was detected by the Northern blot analysis in the pancreas and brain tissue. A nucleotide sequence with high homology with C17 was found in human pancreas cDNA.The sequence was cloned in a plasmid vector for further analysis. Antibodies to chromogranin A were measured by radioimmunoassays using as antigens in vitro transcribed and translated [^<35>S]-methionine-labeled mouse chromogranin A.The prevalence of anti-chromogranin A was approximately 10% in prediabetic female NOD mice. Whereas, the C17 antibody radioassay was not sensitive enough to measure titers of circulating antibodies. We also measured ICA512 (IA-2) antibodies in human subjects with the in vitro transcribed and translated [^<35>S]-methionine-labeled ICA512. The frequency of the ICA512 antibodies was higher in acute-onset IDDM than slowly progressive IDDM.The frequency and titer of ICA512AA declined sharply within 5 years after the onset of IDDM,suggesting the association of the antibody and acute beta-cell destruction.

用糖尿病前期NOD鼠血清免疫筛选NOD小鼠胰岛lambdagt11表达文库，获得两个克隆。克隆(C17和C18)可被NOD鼠血清识别，但不能被Balb/c鼠血清识别。10周龄NOD雌性小鼠血清中C17和C18融合蛋白抗体阳性率分别为80%和10%。C18的DNA序列与小鼠嗜铬粒蛋白A基因(596-1892)完全一致。C17的核苷酸序列显示有一个由46个氨基酸组成的聚(A)尾巴和开放阅读框。Northern印迹法检测胰腺和脑组织中C17mRNA的表达。在人胰腺cDNA中发现了一个与C17高度同源的核苷酸序列，并将其克隆到载体中进行进一步分析。以体外转录和翻译的甲硫氨酸标记的小鼠嗜铬粒蛋白A为抗原，用放射免疫分析法检测了小鼠嗜铬粒蛋白A抗体。然而，C17抗体放射测定法不够灵敏，不能测量循环抗体的滴度。我们还用体外转录和翻译的蛋氨酸标记的ICA512(IA-2)检测了受试者的ICA512(IA-2)抗体。急性起病的IDDM患者ICA512抗体的出现频率高于缓慢进展的IDDM患者，ICA512AA的出现频率和滴度在IDDM发病后5年内急剧下降，提示该抗体与急性胰岛β细胞的破坏有关。

项目成果

Kentaro Yamada,et al.: "Combined Measurements of GAD65 and ICA512 antibodies in acute onset and slowly progressive IDDM" Diabetes Research and Clinical Practice. 印刷中. (1996)

Kentaro Yamada 等人：“GAD65 和 ICA512 抗体在急性发作和缓慢进展的 IDDM 中的联合测量”糖尿病研究和临床实践（1996 年）。

Kentaro Yamada,et al.: "Combined Measurements of GAD65 and ICA512 antibodies in acute onset and slowly progressive IDDM" Diabetes Research and Clinical Practice. (印刷中). (1996)

Kentaro Yamada 等人：“GAD65 和 ICA512 抗体在急性发作和缓慢进展 IDDM 中的联合测量”糖尿病研究和临床实践（1996 年出版）。

Kentaro Yamada, Xiahong Yuan, Chizuko Inada, Hideki Hayashi, Ken-ichi Koyama, Fumi Ichikawa, George S.Eisenbarth, Kyohei Nonaka: "Combined Measurements of GAD65 and ICA512 antibodies in acute onset and slowly progressive IDDM." Diabetes Research and Clini

Kentaro Yamada、Xiahong Yuan、Chizuko Inada、Hideki Hayashi、Ken-ichi Koyama、Fumi Ichikawa、George S.Eisenbarth、Kyohei Nonaka：“GAD65 和 ICA512 抗体在急性发作和缓慢进展 IDDM 中的联合测量。”

Kentaro Yamada, Naoko Takene-Gyotoku, Chizuko Inada, Kyohei Nonaka.: "Endogenous tumor necrosis factor-alpha production by a pancreatic beta-cell line : inhibitory effects of hydrocortisone and nicotinamide." Life Sciences. 59-17. 1423-1429 (1996)

Kentaro Yamada、Naoko Takene-Gyotoku、Chizuko Inada、Kyohei Nonaka.：“胰腺 β 细胞系产生内源性肿瘤坏死因子-α：氢化可的松和烟酰胺的抑制作用。”