Expression of programmed death receptor ligand 1 in B cells and B cell subsets of patients with type 1 diabetes mellitus
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摘要:目的
探讨1型糖尿病(T1DM)患者循环B细胞中程序性死亡受体配体1(PD-L1)的表达情况。
方法采集健康对照者(n=25)和T1DM患者(n=25)的外周血标本。流式细胞术检测B细胞及其表面PD-L1的表达。
结果T1DM患者B细胞亚群频率与健康对照者比较, 差异无统计学意义(P>0.05)。与健康对照者相比, T1DM患者CD19+细胞、CD19+CD27+细胞和CD19+CD27+细胞上PD-L1的表达降低, 差异有统计学意义(P＜0.05)。健康对照者与T1DM患者的PD-L1在B10细胞、边缘区B细胞(MZB)和滤泡B细胞(FoB)上的表达比较，差异无统计学意义(P>0.05)。T1DM患者的PD-L1在过渡性T2-边缘区前体B细胞(T2-MZP)上的表达低于健康对照者，差异有统计学意义(P＜0.05)。
结论PD-L1在T1DM发病中可能起保护作用。PD-L1高表达的B细胞可能为自身免疫性糖尿病患者的治疗提供新策略。
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关键词:
- 1型糖尿病 /
- 程序性死亡受体配体1 /
- B细胞 /
- 流式细胞术
Abstract:ObjectiveTo investigate the expression of programmed death receptor ligand 1 (PD-L1) in circulating B cells of patients with type 1 diabetes mellitus (T1DM).
MethodsPeripheral blood samples were collected from healthy controls (n=25) and T1DM patients (n=25). The expression of PD-L1 on B cells and their surfaces was detected by flow cytometry.
ResultsThere was no significant difference in the frequency of B cell subsets between the T1DM patients and healthy controls (P>0.05). Compared with healthy controls, the expression of PD-L1 on CD19+ cells, CD19+CD27+ cells and CD19+CD27+ cells in the T1DM patients was significantly decreased (P < 0.05). There was no significant difference in the expression of PD-L1 in B10 cells, marginal zone B cells (MZB) and follicular B cells (FoB) between healthy controls and T1DM patients (P>0.05). The expression of PD-L1 on transitional 2-marginal zone precursor B cells (T2-MZP) cells in the T1DM patients was lower than that in the healthy controls (P < 0.05).
ConclusionThe PD-L1 may play a protective role in the pathogenesis of T1DM. B cells with high expression of PD-L1 may provide a new strategy for the treatment of patients with autoimmune diabetes.
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图 1 T1DM和健康对照组受试者的B细胞和B细胞亚群表达
A: B细胞; B: 记忆B细胞; C: 幼稚B细胞; D: B10细胞; E: MZB细胞; F: FoB细胞; G: T2-MZP细胞。
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图 2 CD19+细胞表面PD-L1表达的流式细胞分析
A: CD19+ B细胞的门控策略; B: CD19+细胞中PD-L1表达的门控策略; C: T1DM组与健康对照组CD19+ B细胞上PD-L1的表达比较。每个点代表个体PD-L1的百分比，水平线表示平均值。
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图 3 CD19+CD27+和CD19+CD27-细胞表面PD-L1表达的流式细胞术分析
A: CD19+CD27+和CD19+CD27-细胞中PD-L1表达的门控策略; B: T1DM组与健康对照组CD19+ CD27+细胞上PD-L1的表达比较; C: T1DM组与健康对照组PD-L1在CD19+ CD27-细胞上的表达比较。
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图 4 CD19+CD5+CD1D+细胞表面PD-L1表达的流式细胞术分析
A: CD19+CD5+CD1D+细胞中PD-L1表达的门控策略; B: T1DM组与健康对照组CD19+CD5+CD1D+细胞上PD-L1的表达比较。每个点代表个体PD-L1的比例。水平线表示平均值。
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图 5 MZB、FoB和T2-MZP B细胞上PD-L1表达的流式细胞分析
A: FoB、T2-MZP和MZB细胞中PD-L1的门控策略; B、C、D: T1DM组与健康对照组FoB、T2-MZP和MZB细胞上PD-L1的表达比较。每个点代表个体的PD-L1比例，水平线表示平均值。
表 1 T1DM患者临床特征(n=25)(x±s)[M(IQR)]
临床特征 数值 女性例数 12 胰岛自身抗体阳性例数 18 病程*/年 4.00(6.00) 高密度脂蛋白/(mmol/L) 1.54±0.68 低密度脂蛋白/(mmol/L) 2.61±1.01 胆固醇/(mmol/L) 4.42±1.43 甘油三酯*/(mmol/L) 0.77(0.57) 丙氨酸氨基转移酶/(U/L) 18.28±15.03 天冬氨酸转移酶/(U/L) 17.84±8.15 肌酐/(μmol/L) 56.04±14.19 尿素/(mmol/L) 4.88±1.53 尿酸/(μmol/L) 254.00±97.49 空腹血糖/(mmol/L) 9.80±4.85 空腹c肽*/(ng/mL) 0.08(0.34) 糖化血红蛋白/% 8.28±2.59 尿微量白蛋白肌酐比*/(mg/g) 11.45(12.50) *数据以中位数(四分位数间距)表示。 
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表 2 B细胞亚群PD-L1水平与临床特征之间的相关性
指标 CD19+CD27+ CD19+CD27- B10 MZB FoB T2-MZP 女性 0.061 0.178 0.308 -0.233 -0.033 -0.044 胰岛自身抗体 0.259 0.080 0.178 0.074 0.321 0.012 病程 -0.114 0.196 -0.080 0.078 -0.138 -0.12 高密度脂蛋白 -0.31 0.024 0.287 0.101 -0.23 0.033 4 低密度脂蛋白 -0.062 -0.007 -0.133 -0.124 -0.477 -0.384 胆固醇 -0.042 -0.047 0.021 -0.075 -0.316 -0.163 甘油三酯 -0.011 0.115 0.022 -0.192 -0.325 -0.227 丙氨酸氨基转移酶 -0.017 0.128 0.095 -0.138 -0.296 -0.311 天冬氨酸转移酶 -0.067 0.156 0.11 -0.211 -0.333 -0.317 肌酐 0.097 0.022 -0.17 0.339 0.145 0.019 尿素 0.166 0.129 -0.108 0.619 0.138 0.199 尿酸 0.161 -0.089 -0.045 0.122 0.055 -0.125 空腹血糖 -0.017 -0.068 0.225 -0.188 0.273 0.048 空腹c肽 -0.041 -0.214 -0.212 0.320 0.330 0.214 糖化血红蛋白 -0.093 -0.130 -0.326 -0.259 0.104 -0.149 尿微量白蛋白肌酐比 -0.219 -0.335 0.109 -0.218 -0.120 -0.229 MZB: 边缘区B细胞; FoB: 滤泡B细胞; T2-MZP: 过渡性T2-边缘区前体B细胞。
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