高表达胰岛素反应性天然自身抗体转基因小鼠糖耐量分析  

Glucose Tolerance Analysis on Transgenic Mice with High Titer of Insulin Reactive Natural Autoantibodies

作　　者：宋媛[1] 徐少勇[1] 周洁[1] 高彬[1] 付建芳[1] 邢影[1] 姬秋和[1] 

机构地区：[1]第四军医大学西京医院内分泌代谢科,陕西西安710032

出　　处：《现代生物医学进展》2013年第9期1627-1630,1626,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81070663);国家自然科学基金青年科学基金项目(30901336)

摘　　要：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 determine the glucose tolerance of the transgenic Balb/c and NOD mice with high serum titer of natural autoantibody which reacted with insulin. Methods： ELISA was utilized to detect the polyreactivity of the natural autoantibody in 3B4 tm- nsgenic mice, including insulin and other antigens; 3B4Tg^＋ derived from balb/c background were backeross for five generations to NOD males to produce 3B4Tg^＋/NOD; Oral glucose tolerance test （OGTT） was performed in groups of WT, NOD, 3B4Tg^＋/NOD and 3B4Tg^＋ mice at 10 weeks and 14 weeks of age; Cell suspensions prepared from the peritoneal cavity and spleen were incubated with fluorescent antibody, FACS was employed to detect the development of B cell. Results： The natural autoantibody produced by 3B4 transgenic mice reacted with insulin and other antigens. 3B4Tg^＋/NOD and 3B4Tg^＋mice had abnormal OGTT relative to WT controls at 10 weeks of age and 14 weeks of age. The percentage of B1 a calls in peritoneal B cells and marginal zone B cells in splenic B cells was higher in NOD, 3B4Tg^＋/NOD and 3B4Tg^＋ than WT. Conclusion： Peripheral innate immunity B cells of the natural autoantibody 3B4 transgenic mice differentiate to specific subsets and produce autoantibody, mice producing high level of natural antibody with or without NOD background performance impaired glucose tolerance, these indicate that failure of the innate immunity tolerance is probably one of the pathogenic factors of abnormal glucose metabolism in Type 1 Diabetes mellitus （T1DM）.

关 键 词：3B4天然自身抗体 糖耐量异常 T1DM 天然免疫失耐受 

分 类 号：Q95-3[生物学—动物学] R587.1[医药卫生—内分泌]