急性期川崎病患儿粒细胞样髓源抑制细胞改变及意义初探  

Changes and significance of granulocyte-like myeloid-derived suppressor cells during acute phase of Kawasaki disease

作　　者：温鹏强[1] 王国兵[1] 梅洁花 齐中香[1] 杨莉 徐明国[2] 刘琮[2] 李成荣[1] Wen Pengqiang;Wang Guobing;Mei Jiehua;Qi Zhongxiang;Yang Li;Xu Mingguo;Liu Cong;Li Chengrong(Shenzhen Institute of Pediatrics,Shenzhen Children′s Hospital,Shenzhen 518038,China;Department of Cardiology,Shenzhen Children′s Hospital,Shenzhen 518038,China)

机构地区：[1]深圳市儿童医院儿科研究所,深圳518038 [2]深圳市儿童医院心血管内科,深圳518038

出　　处：《中华微生物学和免疫学杂志》2022年第7期540-548,共9页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(81870364,81102227);深圳市科技计划项目(JCYJ20180228175700233,JCYJ20150403100317055);深圳市医学科研基金(201501032,201401053)。

摘　　要：目的探讨急性期川崎病(Kawasaki disease,KD)患儿粒细胞样髓源抑制细胞(granulocyte-like myeloid-derived suppressor cells,G-MDSC)改变及其在KD免疫发病机制中的作用。方法急性期KD患儿42例,分别于静脉输注丙种球蛋白(intravenous immune globulin,IVIG)治疗前、后直接取血备检,正常同龄儿童32例为对照组。流式细胞术检测外周血HLA-DR-CD11b+CD33+CD14-CD15+G-MDSC比例、活性氧(reactive oxygen species,ROS)浓度以及精氨酸酶-1(arginase-1,Arg-1)、细胞程序性死亡配体1(programmed death-ligand 1,PD-L1)、细胞毒性T淋巴细胞相关蛋白4(cytotoxic T lymphocyte associated protein 4,CTLA4)、糖蛋白130(glycoprotein 130,gp130)、磷酸化信号转导和转录激活因子3(phosphorylated signal transducer and activator of transcription 3,pSTAT3)的蛋白质表达水平;实时荧光定量PCR检测诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、干扰素调节因子8(interferon regulatory factor 8,IRF-8)、IL-6受体α(IL-6 receptorαsubunit,IL-6Rα)、粒细胞集落刺激因子受体(granulocyte colony-stimulating factor receptor,G-CSFR)、CCAAT/增强子强合蛋白β(CCAAT/enhancer binding proteinβ,C/EBPβ)、细胞因子信号抑制物1(suppressor of cytokine signaling 1,SOCS1)、SOCS3 mRNA表达;染色质免疫共沉淀法检测SOCS1、SOCS3基因启动子组蛋白H3乙酰化水平;ELISA检测血浆IL-6、粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)及培养上清中IL-10、转化生长因子-β(transforming growth factor-β,TGF-β)、一氧化氮(nitric oxide,NO)浓度。结果(1)与对照组比较,急性期KD患儿外周血G-MDSC比例、胞内ROS浓度以及Arg-1、PD-L1、CTLA4表达水平明显增高(P<0.05),LPS刺激的G-MDSC培养上清中IL-10、TGF-β浓度亦高于对照组(P<0.05),但合并冠状动脉损伤组(CAL)患儿前述7项指标均低于未合并冠状动脉损伤组(NCAL),差异有统计学意义(P<0.05),经IVIG治疗呈不同程度恢复(P<0.05);各组间iNOS�Objective To investigate the changes and significance of granulocyte-like myeloid-derived suppressor cells(G-MDSC)in the acute phage of Kawasaki disease(KD).Methods Forty-two children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group.The proportion of HLA-DR-CD11b+CD33+CD14-CD15+G-MDSC,the concentration of reactive oxygen species(ROS)and the expression of arginase-1(Arg-1),programmed death-ligand 1(PD-L1),cytotoxic T lymphocyte associated protein 4(CTLA4),glycoprotein 130(gp130)and phosphorylated signal transducer and activator of transcription 3(pSTAT3)at protein level were detected by flow cytometry.Quantitative real-time PCR was used to measure the expression of inducible nitric oxide synthase(iNOS),interferon regulatory factor 8(IRF-8),IL-6 receptorαsubunit(IL-6Rα),granulocyte colony-stimulating factor receptor(G-CSFR),CCAAT/enhancer binding proteinβ(C/EBPβ),suppressor of cytokine signaling 1(SOCS1)and SOCS3 at mRNA level in G-MDSC.Chromatin immunoprecipitation was performed to detect the acetylation of histone H3 at the promoters of SOCS1 and SOCS3 genes.Plasma concentrations of IL-6 and granulocyte colony-stimulating factor(G-CSF)and protein levels of IL-10,transforming growth factor-β(TGF-β)and nitric oxide(NO)in the culture supernatant of G-MDSC stimulated with LPS were measured by ELISA.Results(1)Compared with the control group,the proportion of HLA-DR-CD11b+CD33+CD14-CD15+G-MDSC as well as the concentration of ROS and the expression of inhibitory molecules(Arg-1,PD-L1 and CTLA4)in G-MDSC increased significantly in patients with acute KD(P<0.05).Moreover,the concentrations of IL-10 and TGF-βin culture supernatant of G-MDSC were also higher than those of the control group after stimulation with lipopolysaccharide for 48 h(P<0.05).All of the seven afore-mentioned indexes in KD patients with coronary artery lesion(CAL group)were lower than those in patients without coronary artery lesion(NCAL group)(P<0.05),and restored to some extent

关 键 词：粒细胞样髓源抑制细胞 川崎病 SOCS 组蛋白乙酰化 血管损伤 

分 类 号：R725.4[医药卫生—儿科]