GAD65反应性T细胞检测在成人隐匿性自身免疫糖尿病筛查中的应用研究  

Applications of the detection of GAD65 reactive T cells in latent autoimmune diabetes in adult

作　　者：王婉茹[1] 余靖[1] 孙志纯[1] 王爱红[2] 洪真真 张翼[1] 周智广[3] 

机构地区：[1]福建医科大学附属泉州第一医院,福建泉州362000 [2]解放军第306医院内分泌科,北京100102 [3]中南大学糖尿病中心,湖南长沙410008

出　　处：《中国实用内科杂志》2017年第7期617-620,共4页Chinese Journal of Practical Internal Medicine

基　　金：福建省自然科学基金项目计划(2015J01517);泉州市科技计划项目(2016Z030)

摘　　要：目的探讨酶联免疫斑点试验(ELISPOT)检测谷氨酸脱羧酶反应性分泌IFN-γT细胞(GAD65-T)在成人隐匿性自身免疫糖尿病(LADA)筛查中的应用。方法选择2014年至2016年福建医科大学附属泉州第一医院收治的LADA患者28例,2型糖尿病(T2DM)40例,健康对照20例,放射配体法检测GAD65-Ab,连续密度梯度离心法分离人外周血单个核细胞(PBMC),ELISPOT法检测GAD65-T细胞。将40例T2DM分为GAD65-T细胞ELISPOT检测阳性及阴性组,比较两组的临床特征。结果 GAD65-T细胞数(中位数及95%CI)LADA组为7.2(1.9~11.1)个,T2DM组为3.6(2.9~6.0)个,对照组为0.9(0.2~1.7)个。LADA组的细胞数明显高于T2DM及对照组(P均<0.01);T2DM较对照组具有更高频率的细胞数(P<0.05)。以大于95%CI(对照组)判为阳性,LADA,T2DM及对照组的阳性率分别为71.4%(20/28)、32.5%(13/40)、0。40例T2DM亚组分析显示GAD65-T细胞ELISPOT检测阳性组较阴性组的患者BMI更低、血糖控制更差、空腹C肽水平更低。结论部分GAD65-Ab阴性的T2DM患者存在GAD65-T细胞,联合GAD65-T细胞ELISPOT及GAD65-Ab检测可明显提高LADA的检出率。Objective To explore the application of GAD65 reactive T cells（GAD65-T） detected by enzyme linked immune spot （ELISPOT） in screening LADA patients. Methods Totally 28 LADA patients, 40 T2DM（Type 2 diabetes mellitus） patients and 20 healthy controls were recruited. GAD65-Ab were determined by radioligand assays. Human peripheral blood mononuclear Cells were isolated by Ficoll-Hypaque density gradients. ELISPOT assay was performed to measure the number of spots-forming plaque. One spot-forming represents a GAD65-T cells. The clinical characteristics of two groups（ELISPOT positive or negative） were compared by ELISPOT in 40 T2DM patients. Results Compared with T2DM and control group, the number of GAD65-T cell significantly increased in LADA group （P〈0.01）, there being significantly more GAD65-T cells in T2DM than in control group（P〈0.05）, A positive response was defined when the cell number 〉 95%CI of control subjects. The positive rate was 71.4% （20 of 28） in LADA group, 37.5（ 14 of 40） in T2DM group and 0 in control group. In 40 T2DM patients, the patients in ELISPOT-positive subgroup had lower BMI, higher HbAlc and worse β cells function compared with the negative subgroup. Conclusions GAD65-T cells are found in some patients initially diagnosed with T2DM（GAD65-Ab negative）, combination of enzyme linked immune spot and GAD65-Ab assay can improve the detection rate of LADA.

关 键 词：糖尿病 自身免疫 隐匿性 谷氨酸脱羧酶 T淋巴细胞 酶联免疫斑点 

分 类 号：R587.1[医药卫生—内分泌] R394.6[医药卫生—内科学]