急性期川崎病患儿单核细胞样髓源抑制细胞改变及意义初探  

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

作　　者：俞灵盈 王国兵 温鹏强[2] 梅洁花[3] 齐中香 徐明国[3] 刘琮[3] 李成荣[2] Yu Lingying;Wang Guobing;Wen Pengqiang;Mei Jiehua;Qi Zhongxiang;Xu Mingguo;Liu Cong;Li Chengrong(Zhuhai Campus of Zunyi Medical University,Zhuhai 519041,China;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]遵义医科大学珠海校区,519041 [2]深圳市儿童医院儿科研究所,518038 [3]深圳市儿童医院心血管内科,518038

出　　处：《中华微生物学和免疫学杂志》2021年第10期764-770,共7页Chinese Journal of Microbiology and Immunology

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

摘　　要：目的探讨急性期川崎病(Kawasaki disease,KD)患儿单核细胞样髓源抑制细胞(monocytic myeloid-derived suppressor cells,M-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38例,分别于静脉注射免疫球蛋白(intravenous immunoglobulin,IVIG)治疗前、后直接取血备检,32例正常同龄儿童作为对照组。采用流式细胞术检测外周血HLA-DR-CD11b+CD33+CD15-CD14+M-MDSC和CD4+CD25+CD127-调节性T(Treg)细胞比例、活性氧(reactive oxygen species,ROS)浓度及精氨酸酶1(arginase-1,Arg-1)、CD39、CD73、CD40、CD40L、CCR5蛋白表达水平;实时荧光定量PCR检测诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、细胞毒性T淋巴细胞相关抗原4(cytotoxic T lymphocyte associated antigen-4,CTLA4)、淋巴细胞活化基因3(lymphocyte-activation gene 3,LAG3)mRNA表达水平;ELISA检测培养上清中NO、CCL3、CCL4、CCL5、IL-10、TGF-β浓度。结果(1)与对照组比较,急性期KD患儿外周血M-MDSC比例、胞内ROS浓度及iNOS、CD39、CD73表达水平明显降低(P<0.05),且经LPS刺激后M-MDSC培养上清中NO、IL-10、TGF-β浓度均低于对照组(P<0.05),IVIG治疗后呈不同程度恢复(P<0.05)。各组间Arg-1表达差异无统计学意义(P<0.05)。(2)急性期KD患儿M-MDSC共刺激分子CD40表达显著降低(P<0.05),LPS刺激后M-MDSC培养上清中趋化因子CCL3、CCL4、CCL5浓度均低于对照组(P<0.05),经IVIG治疗后显著上调(P<0.05),但CD40表达仍低于对照组(P<0.05);(3)与对照组比较,急性期KD患儿外周血CD4+CD25+CD127-Treg细胞比例及CTLA4、LAG3、CD40L、CCR5表达水平明显降低(P<0.05),且Treg和M-MDSC比例呈正相关关系(r=0.58,P<0.05),经IVIG治疗后均显著上升(P<0.05)。结论M-MDSC数量不足及功能障碍可能是导致急性期KD患儿免疫功能异常的重要原因。Objective To investigate the changes of monocytic myeloid-derived suppressor cells(M-MDSC)in children with acute Kawasaki disease(KD)and its roles in the immunological pathogenesis of KD.Methods A total of 38 children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group.The proportions of HLA-DR-CD11b+CD33+CD15-CD14+M-MDSC and CD4+CD25+CD127-regulatory T cells(Treg)in peripheral blood,concentrations of reactive oxygen species(ROS)and expression of arginase-1(Arg-1),CD39,CD73,CD40,CD40L and CCR5 at protein levels were detected by flow cytometry.Quantitative real-time PCR was used to evaluate the transcription levels of inducible nitric oxide synthase(iNOS)in M-MDSC and the transcription levels of cytotoxic T-lymphocyte associated antigen 4(CTLA4)and lymphocyte-activation gene 3(LAG3)in Treg.Concentrations of NO,CCL3,CCL4,CCL5,IL-10 and TGF-βin the supernatants of cell culture were measured by ELISA.Results(1)The proportion of HLA-DR-CD11b+CD33+CD15-CD14+M-MDSC,the concentration of intracellular ROS and the expression of iNOS,CD39 and CD73 in M-MDSC decreased significantly in patients with acute KD as compared with those in the control group(P<0.05),and the concentrations of NO,IL-10 and TGF-βin culture supernatant of M-MDSC were lower than those in the control group upon lipopolysaccharide(LPS)stimulation for 48 h(P<0.05).All of the aforementioned indexes restored to some extent after intravenous immunoglobulin(IVIG)therapy(P<0.05).No statistical differences were found in Arg-1 expression between healthy controls and patients with KD before or after IVIG therapy(P<0.05).(2)CD40 expression on M-MDSC was significantly lower in the acute KD group than in the control group(P<0.05).The concentrations of CCL3,CCL4 and CCL5 in the culture supernatants of M-MDSC were lower in the acute KD group than in the control group after LPS stimulation(P<0.05).With IVIG treatment,all of the indexes were up-regulated significantly(P<0.05),although CD40 expression was st

关 键 词：单核细胞样髓源抑制细胞 川崎病 iNOS 共刺激分子 调节性T细胞 

分 类 号：R725.4[医药卫生—儿科]