胰岛素B链基因疫苗的构建及其对小鼠自身免疫性糖尿病的干预预防

Construction of insulin B chain DNA vaccine that prevents mice from autoimmune diabetes

目的 应用构建胰岛素B链基因疫苗预防自身免疫性糖尿病。方法 采用逆转录聚合酶链反应 (RT PCR)等技术 ,构建胰岛素B链基因疫苗 ,将该疫苗注射入胰岛素B链基因疫苗治疗组 (T组 )和胰岛素B链基因疫苗预防组 (P组 )小鼠胫前肌内。定时测定小鼠血糖、胰岛素水平。于第四周观察小鼠胰腺组织病理形态的改变。与对照组 (C组 )和糖尿病组 (D组 )进行比较。结果(1)运用RT PCR扩增技术 ,克隆胰岛素B链基因 ,构建成胰岛素B链基因疫苗。 (2 )在有糖尿病发病倾向小鼠发病前注射胰岛素B链基因疫苗后 ,其发病例数第 2周为 3只 (3/ 10 ) ,第 3、4周为 4只(4/ 10 ) ,显著低于D组和T组 ;P组内预防有效者血中胰岛素水平与C组相比差异无显著性。而糖尿病未缓解的小鼠病理变化和D组相似。结论 构建胰岛素B链基因疫苗可以有效地预防自身免疫性糖尿病的发生 ,为

Objective To construct insulin B chain DNA vaccine and to perform a vaccinating trial for preventing mice from autoimmune diabetes induced by multiple low dose streptozotocin. Methods RT PCR was used to clone insulin B chain, and insulin B chain DNA vaccine was constructed. DNA vaccine was injected into tibilias anterior muscle of C57BL/6 mice in treatment group (T), prophylactic group (P). The morphology of mice islets was investigated by pathology, blood samples were taken to determine glucose and insulin. Group T and group P were compared with group of normal control mice (C) and group of untreated diabetic mice (D). Results (1) Insulin B chain gene was cloned by RT PCR, which ligated with pcDNA3 plasmid. Insulin B chain DNA vaccine was constructed. (2) In group P, after injection of DNA vaccines, 3 (3/10) and 4 (4/10) mice became diabetic in the second week and the third, fourth week respectively, the incidence was significantly less than those of groups D and T. The insulin level showed no difference between group C and the non diabetic mice of group P. The change of pancreas of diabetic mice in group P was similar to group D. Conclusion Insulin B chain DNA vaccine can prevent pre autoimmune diabetes.