1型糖尿病患者外周血新型CD19^(+)CD27^(+)CD39^(hi)调节性B细胞的鉴定与分析  

Identification and analysis of novel CD19+CD27+CD39hi regulatory B cells in the peripheral blood of patients with type 1 diabetes mellitus

作　　者：秦瑶[1] 李雨潇 屈馨怡 魏倩影 陈欣[1] 邵瀛 王苏雨 王星[1] 张梅[1] Qin Yao;Li Yuxiao;Qu Xinyi;Wei Qianying;Chen Xin;Shao Ying;Wang Suyu;Wang Xing;Zhang Mei(Department of Endocrinology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院内分泌科,南京210029

出　　处：《中华糖尿病杂志》2024年第7期783-789,共7页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家自然科学基金(82000747,82370828);中华国际医学交流基金会医路恒睿糖尿病代谢科研基金(Z-2017-26-2202-4);江苏省医学重点学科(ZDXK202202)。

摘　　要：目的探讨CD19+CD27+CD39hi调节性B细胞(Breg)在1型糖尿病(T1DM)患者中的特征及临床意义。方法为病例对照研究。招募自2022年1至8月在南京医科大学第一附属医院就诊的49例T1DM患者和20名健康对照者,采集研究对象的一般信息和病史,检测T1DM患者的空腹血糖、糖化血红蛋白(HbA1c)、血清胰岛素和C肽、糖尿病胰岛自身抗体等临床指标;采用流式细胞术分析CD19+B细胞、CD19+CD27+CD39hi Breg比例及白细胞介素(IL)-10、程序性死亡配体1(PD-L1)的表达;通过酶联免疫吸附法测定血清IL-1β、IL-6和肿瘤坏死因子-α(TNF-α)的水平。组间比较采用独立样本t检验、Mann-WhitneyU检验或χ^(2)检验。采用Spearman相关分析法评估Breg比例与炎性细胞因子水平之间的相关性。结果与健康对照者相比,T1DM患者外周血CD19+B细胞比例没有变化,CD19+CD27+CD39hi Breg比例显著增加[分别为3.45%(1.80%,7.96%)和1.28%(1.04%,2.17%),P<0.001],但其胞内IL-10的表达水平下降(分别为4.96%±3.15%和9.47%±6.52%,P=0.007),PD-L1表达无明显改变。进一步分析发现,发病年龄≥18岁[分别为6.42%(2.89%,11.53%)和2.37%(1.42%,5.26%),P=0.004]、病程≤1年[分别为5.74%(2.60%,12.58%)和2.95%(1.47%,5.83%),P=0.026]或峰值C肽>3.30 pmol/L[分别为5.27%(2.19%,10.38%)和2.39%(1.14%,5.42%),P=0.029]的患者CD19+CD27+CD39hi Breg比例更高。Spearman相关分析结果显示,Breg比例与血清IL-1β水平呈正相关(r=0.360,P=0.011)。然而,Breg内IL-10的表达与患者的临床特征及炎性细胞因子水平均无关(P>0.05)。结论 T1DM患者外周血CD19+CD27+CD39hi Breg数量及功能存在异常,本研究结果进一步补充了T1DM免疫失衡的临床证据。Objective To investigate the characteristics and clinical significance of CD19+CD27+CD39hi regulatory B cells(Breg)in patients with type 1 diabetes mellitus(T1DM).Methods This was a case-control study.A total of 49 T1DM patients and 20 healthy controls were recruited from the First Affiliated Hospital of Nanjing Medical University from January to August 2022.General information and medical history of the subjects were collected.Fasting plasma glucose,glycated hemoglobin A1c(HbA1c),serum insulin and C-peptide,islet autoantibodies and other clinical indicators were measured in T1DM patients.The proportion of CD19+B cells,CD19+CD27+CD39hi Breg and the expression of interleukin(IL)-10 and programmed death ligand 1(PD-L1)were analyzed by flow cytometry.Serum levels of IL-1β,IL-6 and tumor necrosis factor-α(TNF-α)were determined by enzyme-linked immunosorbent assay.Independent samples t-test,Mann-Whitney U test orχ^(2) test was used for comparison between groups.Spearman correlation analysis was used to evaluate the correlation between the proportion of Breg and inflammatory cytokine levels.Results Compared with healthy controls,the proportion of total CD19+B cells in peripheral blood of T1DM patients did not change,the proportion of CD19+CD27+CD39hi Breg increased significantly[3.45%(1.80%,7.96%)vs.1.28%(1.04%,2.17%),P<0.001],but the expression level of intracellular IL-10 decreased(4.96%±3.15%vs.9.47%±6.52%,P=0.007),and the expression of PD-L1 did not change significantly.Further analysis showed that the proportion of CD19+CD27+CD39hi Breg was higher in patients with age of onset≥18 years[6.42%(2.89%,11.53%)vs.2.37%(1.42%,5.26%),P=0.004],duration≤1 year[5.74%(2.60%,12.58%)vs.2.95%(1.47%,5.83%),P=0.026]or peak C-peptide>3.30 pmol/L[5.27%(2.19%,10.38%)vs.2.39%(1.14%,5.42%),P=0.029],and was positively correlated with serum IL-1βlevels(r=0.360,P=0.011).However,intracellular IL-10 expression in Breg was not associated with clinical features or inflammatory cytokine levels(P>0.05).Conclusion The number and funct

关 键 词：糖尿病 1型 白细胞介素-10 调节性B细胞 程序性死亡配体1 

分 类 号：R587.1[医药卫生—内分泌]