ELISPOT检测羧基肽酶H阳性成人隐匿性自身免疫性糖尿病患者羧基肽酶H反应性T细胞  

Detection of carboxypeptidase H specific T cells in peripheral blood of latent autoimmune diabetic patients with carboxypeptidase antibody positivity by ELISPOT assay

作　　者：杨琳[1] 周智广[1] 杜弢[1] 谭少珍[1] 张翼[1] 金萍[1] 

机构地区：[1]中南大学湘雅二医院内分泌科,中南大学代谢内分泌研究所,中南大学糖尿病中心,长沙410011

出　　处：《中南大学学报（医学版）》2009年第10期1011-1016,共6页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(30400217);教育部博士点专项科研基金(20030533033)~~

摘　　要：目的:探讨羧基肽酶H抗体(CPH-Ab)阳性的成人隐匿性自身免疫性糖尿病(LADA)患者外周血T细胞免疫的特点。方法:选取CPH-Ab+LADA患者42例,2型糖尿病组(T2DM)患者20例和健康对照22例,分离外周血单个核细胞;自制人CPH蛋白,应用酶联免疫斑点(ELISPOT)技术检测并比较各组在CPH诱导下分泌IFN-γ的Th1和分泌IL-4的Th2细胞数,计算Th1与Th2比值(Th1/Th2)。结果:与健康对照和T2DM比较,CPH-Ab+LADA患者的IFN-γ-T和IL-4-T细胞数无异常增多,Th1/Th2亦无统计学差异(P>0.05);但22例短病程(<3年)CPH-Ab+患者的CPH-IL-4-Th2细胞数较T2DM患者和正常对照均显著升高(1.8 vs.0.2和0.3,均P<0.05),但未发现任何因素与该斑点数相关。各组间的T细胞针对植物血凝素的反应均无统计学差异。结论:CPH未直接诱导LADA的细胞免疫破坏反应,CPH反应性T细胞标志的可能是一进展相对缓慢的、胰岛β细胞的自身免疫过程。Objective To explore the characteristics of T cell immunity in peripheral blood of patients with carboxypeptidase-H antibody（CPH-Ab）.Methods Forty-two latent autoimmune diabetes in adults（LADA） patients with CPH-Ab^＋ alone,20 Type 2 diabetes mellitus patients（T2DM）,and 22 healthy controls were selected and their peripheral blood mononuclear cells were isolated.Human recombinant carboxypeptidase（CPH） protein was expressed and further used as a stimulant in Enzyme-linked immunospot（ELISPOT） assay to detect IFN-γ-Th1 and IL-4-Th2 cells in the 3 groups.Th1/Th2 ratios were also calculated.CPH-Ab and glutamic acid decarboxylase antibody（GAD-Ab） were determined by radioligand assay.Results Compared with healthy controls and T2DM,IFN-γ-Th1 and IL-4-Th2 numbers did not increase significantly in CPH-Ab^＋ group,nor did the Th1/Th2 ratios（P〉0.05）.We further divided the CPH-Ab^＋ patients into a short duration group（n=22） and a long duration subgroup（n=20） according to the duration of 3 years.CPH-IL-4-T in the short duration subgroup was significantly higher than that in T2DM and healthy controls（1.8 vs.0.2 and 0.3,both P〈0.05） and we did not find any factor that was significantly correlated with the IL-4 spots number.There were not any significant differences in T cell responses to phaseolus vulgaris agglutinin（PHA） among all groups（P〉0.05）.Conclusion CPH does not directly involve in the cellular pathological mechanism of LADA.Anti-CPH immunity may be associated with more slowly aggressive beta cell autoimmunity.

关 键 词：羧基肽酶H 成人隐匿性自身免疫性糖尿病 酶联免疫斑点 Th1/Th2型淋巴细胞 

分 类 号：R587.1[医药卫生—内分泌]