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目的探讨成人隐匿性自身免疫糖尿病(LADA)患者外周血单个核细胞(PBMC)对胰岛素和谷氨酸脱羧酶(GAD)的增殖反应。方法密度梯度离心法分离21例LADA、25例2型糖尿病(DM)及20名正常对照PBMC,3HTdR掺入法检测其对胰岛素和GAD及其分别联合白细胞介素2(IL2)刺激的增殖反应,以刺激指数(SI)反映增殖反应强度。结果(1)PBMC对PHA、胰岛素或GAD刺激的增殖反应在LADA、2型DM和正常对照组间差异无统计学意义。(2)对于胰岛素或GAD联合IL2(胰岛素+IL2,GAD+IL2)刺激,LADA组SI高于2型DM组(5.1vs0.8;;7.0vs0.8,均P<0.05);;胰岛素+IL2刺激较胰岛素或IL2单独刺激的倍增值,GAD+IL2刺激较GAD或IL2单独刺激的倍增值LADA组高于2型DM组(4.0vs1.0;;3.4vs0.3;;6.3vs1.2;;4.6vs0.3,均P<0.05)。(3)LADA患者胰岛素刺激的SI胰岛素与GAD抗体指数呈负相关。结论LADA患者体内存在已致敏的T细胞,其活化效应在特异性抗原(GAD或胰岛素)和IL2共刺激时方可被发现。
Objective To investigate the proliferative responses of peripheral blood mononuclear cells (PBMCs) to insulin and glutamic acid decarboxylase (GAD) in adults with latent autoimmune diabetes (LADA). Methods 21 cases of LADA, 25 cases of type 2 diabetes mellitus (DM) and 20 normal control PBMCs were isolated by density gradient centrifugation. Proliferation response to insulin and GAD and their combination with interleukin-2 (IL2) stimulation was detected by 3H-thymidine incorporation , Stimulated index (SI) reflect the proliferation reaction intensity. Results (1) There was no significant difference in the proliferative response of PBMC to PHA, insulin or GAD stimulation between LADA, type 2 DM and normal control group. (2) SI in LADA group was higher than that in 2 DM group (5.1vs0.8 ;; 7.0vs0.8, all P <0.05) for insulin or GAD combined with IL2 (insulin + IL2, GAD + IL2) IL2 stimulation than insulin or IL2 alone stimulated doubling value, GAD + IL2 stimulation than GAD or IL2 alone stimulated double the value of LADA group was higher than the type 2 DM group (4.0vs1.0 ;; 3.4vs0.3 ;; 6.3vs1.2 ;; 4.6vs0.3, all P <0.05). (3) Insulin-stimulated SI insulin in LADA patients was negatively correlated with GAD antibody index. Conclusions Sensitized T cells exist in LADA patients. The activation of T cells can be detected only when specific antigen (GAD or insulin) and IL2 are costimulated.