TLR9联合CD40信号对ITP患儿外周血中IL10^+CD3^-细胞亚群改变的研究  

The role of TLR9 and CD40 signaling axis in regulation of IL10^+ CD3^- subsets from PBMC of children ITP patients

作　　者：郑蒙蒙 施建栋 周海霞[2] ZHENG Mengmeng;SHI Jiandong;ZHOU Haixia(Flow Cytometry Core Facility,Scientific Research Center,Wenzhou Medical University,Wenzhou 325035,China;Department of Hematology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学科研实验中心流式细胞技术平台,浙江温州325035 [2]温州医科大学附属第二医院育英儿童医院儿童血液科,浙江温州325027

出　　处：《温州医科大学学报》2020年第7期568-572,共5页Journal of Wenzhou Medical University

基　　金：浙江省公益技术应用研究(实验动物)项目(2016C37113);温州市科技计划项目(Y20150024);浙江省医药卫生科技计划项目(2009A148)。

摘　　要：目的:探讨TLR9联合CD40信号对儿童免疫性血小板减少症(ITP)患儿外周血单个核细胞(PBMC中CD3^-细胞表达IL-10的作用及影响。方法:采用流式多因子阵列检测技术,分析8例ITP患儿和7例正常对照血浆样本中IL-10含量;分离8例ITP患儿及7例正常对照外周血PBMC,并经CpG联合CD40L体外刺激培养5 h,采用流式胞内外染色及检测技术,比较ITP患儿与正常对照组PBMC中CD3^-和CD3^+亚群比例、IL-10表达能力及细胞存活率的差异。结果:ITP患儿血浆中IL-10含量显著高于正常对照组(P<0.05);CpG联合CD40L刺激后的ITP患儿和正常对照组PBMC中CD3^-细胞比例与未刺激组相比均显著升高,而CD3^+T淋巴细胞比例显著下降(P<0.05);ITP患儿刺激组中CD3^-IL-10^+细胞的比例显著高于正常对照刺激组(P<0.05);与正常对照组相比,未刺激组ITP患儿CD3^-细胞存活率显著下降(P<0.05)。结论:TLR9联合CD40信号的活化能导致ITP患儿外周血IL10^+CD3^-细胞比例升高,细胞死亡率升高,可能与ITP疾病进展有重要关系。Objective:To study the effects of TLR9 combined with CD40 signal on the CD3^-population and IL-10 production in the PBMCs from children with ITP.Methods:The plasma IL-10 level in 8 ITP children and healthy controls were determined by using flow cytometry based LEGENplex panel technology.PBMCs from 8 ITP children and 7 healthy controls were isolated by using human lymphocytes separation buffer,and subjected to CpG plus CD40 L stimulation invitro for 5 hours.Then cells were analyzed by flow cytometry after staining with the indicated markers.Results:Compared with controls(5.677±0.808),elevated plasma IL-10 level(8.063±0.892)was observed in ITP children.The percentage of CD3^-cells in PBMCs was significantly increased in both CpG plus CD40 L stimulation and unstimulated group.However,percentage of CD3^+cells was decreased after stimulation.Moreover,the IL-10 production was robustly enhanced in stimulated PBMCs from ITP children.In addition,compared with unstimulated control samples,the unstimulated PBMCs samples from ITP children showed reduction in cell viability.Conclusion:The TLR9 signal together with CD40 signal is crucial for triggering both IL-10 production and cell-death in CD3^-cells from children with ITP.

关 键 词：TOLL样受体9 CD40配体 ITP 儿童 CD3阴性 白细胞介素10 

分 类 号：R392[医药卫生—免疫学]