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目的:初步探讨高表达胰岛素反应性天然自身抗体对野生及NOD小鼠糖耐量的影响。方法:酶联免疫吸附法(ELISA)检测3B4天然自身抗体与胰岛素及其它自身抗原识别情况;杂交NOD与3B4Tg+小鼠并回交5代以上得到3B4Tg+/NOD小鼠;对不同鼠龄WT、NOD、3B4Tg+/NOD和3B4Tg+小鼠进行口服糖耐量试验(OGTT);获取小鼠脾细胞和腹腔细胞,荧光抗体染色后用流式细胞仪分析不同基因型小鼠B细胞发育耐受情况。结果:3B4天然自身抗体可以识别包括胰岛素在内的多种抗原;10周、14周OGTT时3B4Tg+/NOD、3B4Tg+血糖较WT对照组均显著升高;与WT小鼠相比,NOD、3B4Tg+/NOD和3B4Tg+小鼠外周成熟B细胞明显向腹腔B1a及脾边缘带(MZ)B细胞分化。结论:天然自身抗体3B4转基因小鼠中天然免疫B细胞可以向特定亚群发育分化成熟并分泌自身抗体,高表达天然自身抗体小鼠无论是否具有NOD基因背景均可出现糖耐量异常,提示天然免疫失耐受可能是1型糖尿病糖代谢异常的发病因素之一。
Objective: To investigate the effects of high expression of natural insulin-reactive autoantibodies on glucose tolerance in wild and NOD mice. Methods: The natural autoantibodies of 3B4 and the identification of insulin and other autoantigens were detected by enzyme-linked immunosorbent assay (ELISA); 3B4Tg + / NOD mice were obtained by hybridization of NOD with 3B4Tg + mouse and back for more than 5 generations; , 3B4Tg + / NOD and 3B4Tg + mice. The spleen cells and peritoneal cells were harvested. Fluorescent antibody was used to analyze the developmental tolerance of B cells in different genotypes by flow cytometry. Results: Compared with WT mice, 3B4Tg + / NOD and 3B4Tg + blood glucose at 3 weeks and 14 weeks of OGTT significantly increased compared with WT mice, The peripheral mature B cells of NOD and 3B4Tg + mice differentiated into peritoneal B1a and splenic border zone (MZ) B cells. CONCLUSIONS: Natural B cells from innate autoantibody 3B4 mice can differentiate into mature and secrete autoantibodies to specific subpopulations. Highly expressed natural autoantibodies may show impaired glucose tolerance in NOD-deficient mice, suggesting that innate immunity Intolerance may be one of the causative agents of abnormal glucose metabolism in type 1 diabetes.