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目的:探讨长病程1型糖尿病(T1DM)患者残余胰岛功能与自身免疫状态的关系。方法:采用放射配体法测定的血清C肽对中南大学湘雅二医院T1DM综合管理门诊中长病程(病程≥10年)自身免疫T1DM患者的胰岛功能进行评价。将空腹或餐后2 h C肽高于仪器检测敏感度下限(16.7 pmol/L)定义为胰岛功能存留,否则为胰岛功能丧失。筛查并入组所有胰岛功能存留患者(19例),同时入组性别、年龄、病程、体质指数(BMI)匹配的胰岛功能丧失患者(19例)及健康对照(19例)。检测研究对象外周血CD4+T淋巴细胞亚群:Th1辅助细胞(Th1)、Th2辅助细胞(Th2)、Th17辅助细胞(Th17)、调节性T细胞(Treg)及B淋巴细胞亚群:边缘区B细胞(MZB)、滤泡状B细胞(FoB)、分泌白细胞介素10的调节性B细胞(B10)频率及CD4+T、CD8+T、CD19+B细胞表面程序性死亡受体1(PD-1)及其配体(PD-L1)的表达水平;测定研究对象空腹血清中炎症因子干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-1受体抗体(IL-1RA)、白细胞介素-4(IL-4)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-12p40亚基(IL-12p40)、白细胞介素-12p70亚基(IL-12p70)、白细胞介素-23(IL-23)及干扰素诱生蛋白-10(IP-10)的水平。比较上述指标在胰岛功能存留组、胰岛功能丧失组患者与健康对照之间的差异。结果:19例胰岛功能存留组患者中男性占42.1%(8例),年龄[n M(n Q1,n Q3)]为29.0(23.0,40.0)岁,病程[n M(n Q1,n Q3)]为11.0(10.0,14.0)年;19例胰岛功能丧失组患者中男性占42.1%(8例),年龄[n M(n Q1,n Q3)]为30.0(23.0,40.0)岁,病程[n M(n Q1,n Q3)]为12.0(10.0,15.0)年;19例健康对照中男性占42.1%(8例),年龄[n M(n Q1,n Q3)]为29.0(24.0,40.0)岁。健康对照、胰岛功能丧失组、胰岛功能存留组患者Th1细胞频率[n M(n Q1,n Q3)]分别为9.93%(7.45%,15.20%)、14.90%(11.70%,18.00%)、10.20%(6.93%,15.80%)(n P=0.015);Treg细胞频率[n M(n Q1,n Q3)]分别为3.52%(2.92%,5.68%)、2.88%(1.64%,3.22%)、3.12%(2.81%,4.81%)(n P=0.005);B淋巴细胞表面PD-1分子表达比例[n M(n Q1,n Q3)]分别为4.69%(2.64%,6.37%)、2.11%(1.45%,3.63%)、4.20%(2.53%,6.01%)(n P=0.003);血清IL-6水平[n M(n Q1,n Q3)]分别为26.43(18.06,33.35)ng/L、42.97(25.52,66.30)ng/L、22.07(14.85,34.45)ng/L(n P=0.006);血清IP-10水平[n M(n Q1,n Q3)]分别为107.39(76.19,126.07)ng/L、188.82(131.27,348.18)ng/L、128.26(114.31,136.50)ng/L(n P<0.001)。两两比较发现,胰岛功能丧失组Th1细胞频率、血清 IL-6、IP-10水平均分别高于胰岛功能存留组及健康对照,Treg细胞频率、B 淋巴细胞表面 PD-1 分子表达比例均分别低于胰岛功能存留组及健康对照(均n P<0.05)。n 结论:胰岛功能存留的长病程T1DM患者外周血Th1细胞频率、IL-6、IP-10因子水平更低,Treg细胞频率、B细胞表面PD-1分子水平更高,提示存在更强的自身免疫耐受。“,”Objective:We aimed to investigate the autoimmune status of long-term type 1 diabetes mellitus (T1DM) patients with residual β-cell function.Methods:The residual β-cell function of long-term (disease duration≥10 years) autoimmune T1DM patients from the T1DM Integrated Management Clinic of the Second Xiangya Hospital was assessed by serum C-peptide levels. Patients with fasting or 2-hour postprandial C-peptide levels over the lower sensitivity limit of detection (16.7 pmol/L) were grouped as C-peptide-positive, and others were grouped as C-peptide-negative. We screened and enrolled all the C-peptide-positive patients (n n=19). C-peptide-negative patients with matched sex, age, duration, BMI (n n=19) and healthy controls (n n=19) were recruited at the same time. The frequencies of CD4+T cell (Th1/Th2/Th17/Treg) and B cell (MZB/FoB/B10) subsets, the expression of PD-1/PD-L1 on T and B lymphocytes, and the levels of T1DM related cytokines including IFN-γ, TNF-α, IL-1β, IL-1RA, IL-4, IL-6, IL-10, IL-12p40, IL-12p70, IL-23 and IP-10 were tested. We compared these parameters in patients with different levels of β-cell function.n Results:In healthy controls, C-peptide-negative and C-peptide-positive patients, the frequencies n M (n Q1, n Q3) of Th1 cells were 9.93% (7.45%, 15.20%), 14.90% (11.70%, 18.00%) and 10.20% (6.93%, 15.80%) (n P=0.015), and the frequencies n M (n Q1, n Q3) of Treg cells were 3.52% (2.92%, 5.68%),2.88% (1.64%, 3.22%) and 3.12% (2.81%, 4.81%) (n P=0.005), and the frequencies n M(n Q1,Q3) of PD-1+B cells were 4.69% (2.64%, 6.37%), 2.11% (1.45%, 3.63%) and 4.20% (2.53%, 6.01%) (n P=0.003), respectively. The levels of IL-6 n M(n Q1,n Q3)were 26.43(18.06, 33.35) ng/L, 42.97 (25.52, 66.30) ng/L, and 22.07 (14.85, 34.45) ng/L (n P=0.006), and the levels of IP-10 n M(n Q1,n Q3) were 107.39 (76.19, 126.07) ng/L, 188.82 (131.27, 348.18) ng/L and 128.26 (114.31, 136.50) ng/L (n P<0.001) in healthy controls, C-peptide-negative and C-peptide-positive patients, respectively. Compared with C-peptide-positive patients, the frequency of Th1 cells and the levels of IL-6 and IP-10 cytokines were higher, while the frequencies of Treg cells and PD-1+B cells were lower in C-peptide-negative patients (alln P<0.05).n Conclusions:Long-term T1DM patients with residual β-cell function had lower frequency of Th1 cells, lower levels of IL-6, IP-10 cytokines, and higher frequencies of Treg and PD-1+B cells, which indicated a pronounced autoimmune tolerance.