基于谷氨酸脱羧酶65的口服疫苗对链脲佐菌素诱导的1型糖尿病小鼠的治疗作用  

Therapeutic effect of oral vaccine based on glutamate decarboxylase 65 on streptozotocin-induced type 1 diabetic mice

作　　者：吴伊玲 尤琪 吴洁[1] WU Yiling;YOU Qi;WU Jie(School of Life Science and Technology,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国药科大学生命科学与技术学院,南京211198

出　　处：《中国药科大学学报》2021年第5期614-621,共8页Journal of China Pharmaceutical University

摘　　要：探讨基于谷氨酸脱羧酶65(GAD65)的口服疫苗对链脲佐菌素(STZ)诱导的1型糖尿病(T1D)模型小鼠的治疗作用。采用小剂量多次腹腔注射STZ建立T1D模型。使用溶剂扩散法制备融合蛋白CTB-GADIII与海藻酸钙纳米疫苗(Ca-Alg-GADIII),灌胃T1D小鼠,1周1次,连续5周,每周记录小鼠血糖和体重。通过糖耐量测定(OGTT)和胰腺组织病理分析研究口服疫苗抗T1D药效学。通过ELISA检测血清中谷氨酸脱羧酶抗体(GADA)和胰岛素自身抗体(IAA)、相关细胞因子IL-4、IFN-γ和TGF-β1的含量,流式检测CD4^(+)T细胞分型情况,对口服疫苗抗T1D作用的免疫学机制进行初步探讨。结果显示,疫苗免疫后的小鼠空腹血糖有一定的改善,糖耐量提高,胰腺损伤减少,胰岛素分泌增加,GADA及IAA滴度显著下降,肠系膜淋巴结和胰腺淋巴结中CD4^(+)T细胞免疫平衡有一定的改善。实验结果表明Ca-Alg-GADIII口服疫苗对STZ诱导的T1D小鼠具有一定的治疗作用。To investigate the therapeutic effect of oral vaccine based on glutamate decarboxylase 65(GAD65)on streptozotocin(STZ)-induced type 1 diabetic(T1D)mice,the mice model of T1D was established by intraperitoneal injection of low dose multiple STZ.CTB-GADIII encapsulated with calcium alginate(Ca-Alg-GADIII)was formulated using crosslinking technology with sodium alginate and calcium chloride,and was administered intragastric to T1D mice once a week for 5 consecutive weeks.Blood glucose and body weight of the mice were recorded weekly,and pharmacodynamics against T1D of Ca-Alg-GADIII were investigated by glucose tolerance assay(OGTT)and pancreatic histopathological analysis.The levels of glutamic acid decarboxylase antibody(GADA),and insulin autoantibody(IAA)and related cytokines(IL-4,IFN-γ,TGF-β1)in serum were detected by ELISA,and the CD4^(+)T cell subsets were detected by flow cytometry.The immunological mechanism of oral vaccine against T1D was preliminarily discussed.The results showed that the disease-related indicators improved in immunized mice:fasting blood glucose improved,glucose tolerance and insulin secretion increased,pancreatic injury decreased,autoantibodies like GADA and IAA titers significantly decreased,and CD4^(+)T cell immune balance in mesenteric lymph node(MLN)and pancreatic lymph node(PLN)improved to some extent.The results suggest that oral vaccine Ca-Alg-GADIII has some therapeutic effect on STZ-induced T1D mice.

关 键 词：1型糖尿病 谷氨酸脱羧酶65 海藻酸钙 口服疫苗 免疫耐受 

分 类 号：R967[医药卫生—药理学]