阻塞性睡眠呼吸暂停综合征患者髓源性抑制细胞的免疫功能及其机制  

Immune function of myeloid-derived suppressor cells and its mechanism in obstructive sleep apnea syndrome

作　　者：陈思文[1] 李洁[1] 向彬 徐珅杰 王翎[1] Chen Siwen;Li Jie;Xiang Bin;Xu Shenjie;Wang Ling(Department of General Medical Ward,First Affiliated Hospital,Soochow University,Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院全科医学科,苏州215006

出　　处：《中华医学杂志》2020年第4期295-300,共6页National Medical Journal of China

基　　金：国家自然科学基金青年基金(81500068);江苏省青年医学重点人才(QNRC2016743)。

摘　　要：目的探讨阻塞性睡眠呼吸暂停综合征(OSAS)患者髓源性抑制细胞(MDSC)的免疫功能及其机制。方法选取2015年1月至2016年12月就诊于苏州大学附属第一医院睡眠中心经多导睡眠监测确诊的呼吸暂停低通气指数(AHI)>30次/h的OSAS患者20例(OSAS组)和年龄匹配的健康体检者(健康对照组)20名,采用流式细胞术检测其外周血单核样CD14+低表达或者缺乏人类白细胞DR抗原(HLA-DR-/low)MDSC占CD14+单核细胞的比例。流式细胞仪分选两组外周血CD14+HLA-DR-/low MDSC,磁珠阴性选择法分选人外周血中的T细胞;T细胞增殖试验中,将经羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记的人T细胞与自体MDSC共孵育,流式细胞仪检测各组T细胞的CFSE荧光强度。酶联免疫吸附(ELISA)法和Western印迹法检测两组MDSC中白细胞介素(IL)-6、肿瘤坏死因子α(TNF-α)、IL-10、转化生长因子β1(TGF-β1)的浓度和程序性死亡配体1(PD-L1)的表达阳性率及精氨酸酶1(Arg1)、诱导型一氧化氮合酶(iNOS)、缺氧诱导因子-1α(HIF-1α)的相对表达量。结果OSAS组外周血CD14+HLA-DR-/low MDSC占CD14+单核细胞的比例显著高于健康对照组[(12.5±1.5)%比(3.5±0.4)%,P<0.05];OSAS组MDSC抑制T细胞增殖的能力显著强于健康对照组MDSC[(23.2±1.1)%比(53.7±3.2)%,P<0.05];OSAS组MDSC中IL-6、TNF-α、IL-10、TGF-β1的浓度均显著高于健康对照组[(1316±163)比(642±72)ng/L、(316±35)比(167±18)ng/L、(385±42)比(108±26)ng/L、(44276±4589)比(9557±1124)ng/L](均P<0.05);OSAS组MDSC的膜分子PD-L1表达阳性率显著高于健康对照组[(75.6±7.9)%比(30.6±2.5)%,P<0.05];OSAS组MDSC中Arg1、iNOS、HIF-1α相对表达量分别为健康对照组的(4.6±0.5)、(2.8±0.3)、(4.3±0.4)倍(均P<0.05)。结论OSAS患者可能通过活化HIF-1α信号等诱导MDSC数量增加、免疫抑制相关分子表达升高,进而增强MDSC的免疫抑制功能。Objective To investigate the immune function of myeloid-derived suppressor cells(MDSC)and its mechanism in obstructive sleep apnea syndrome(OSAS).Methods Twenty OSAS patients who were diagnosed by polysomnography(Apnea Hyponea Index>30 events/h)from Sleep Disorders Center at First Affiliated Hospital between January 2015 and December 2016 were selected.The percent of CD14+low expression or lack of human leukocyte antigen DR(HLA-DR-/low)MDSC in the CD14+monocyte from both OSAS patients and healthy people were analyzed by flow cytometry.In vitro assay,MDSC from OSAS patients and health people were sorted by flow cytometry and T cells were sorted with negative isolation kit.For T-cell proliferation assays,the carboxyfluorescein diacetate succinimidyl ester(CFSE)-labeled T cells were respectively incubated with autologous MDSC.CFSE fluorescence intensity of T cells was detected by flow cytometry.Enzyme-linked immunosorbent assay(ELISA)and Western blot analysis were used to evaluate the concentrations of interleukin(IL)-6,tumor necrosis factor-α(TNF-α),IL-10,transforming growth factor-β1(TGF-β1),positive rate of programmed death ligand-L1(PD-L1),relative transcript level of Arginase 1(Arg1),inducible nitric oxide synthase(iNOS),hypoxic inducible factor-1α(HIF-1α)expressed by MDSC.Results Comparing to healthy people,the percentage of CD14+HLA-DR-/low MDSC in CD14+monocyte was significantly elevated[(12.5±1.5)%vs(3.5±0.4)%,P<0.05].In vitro,OSAS patient-derived MDSC exhibited a stronger suppressive effect on T-cell proliferation[(23.2±1.1)%vs(53.7±3.2)%,P<0.05].Further analysis revealed that OSAS patient-derived MDSC secreted much higher concentrations of IL-6,TNF-α,IL-10 and TGF-β1[(1316±163)vs(642±72)ng/L,(316±35)vs(167±18)ng/L,(385±42)vs(108±26)ng/L and(44276±4589)vs(9557±1124)ng/L](all P<0.05).The percentage of membrane molecule PD-L1-positive cells in OSAS patient-derived MDSC was obviously higher than that in healthy people-derived MDSC[(75.6±7.9)%vs(30.6±2.5)%,P<0.05].Compared with healthy pe

关 键 词：睡眠呼吸暂停 阻塞性 抑制细胞 骨髓源性 免疫 

分 类 号：R76[医药卫生—耳鼻咽喉科]