ITP患者骨髓间质干细胞体外对CD4(+)CD25(+)Treg细胞增殖的影响  

Effect of mesenchymal stem cells on CD4(+)CD25(+) Treg cells proliferation in patients with ITP in vitro

作　　者：余先球[1] 陈红梅[1] 孙晋华 罗鸣[1] 陆益龙[1] 

机构地区：[1]江苏大学附属医院血液科,江苏镇江2120012 [2]厦门长庚医院ICU科

出　　处：《临床血液学杂志》2015年第1期26-28,31,共4页Journal of Clinical Hematology

基　　金：镇江市科技支撑计划(社会发展)(No:FZ2011045)

摘　　要：目的:体外研究免疫性血小板减少症(ITP)患者骨髓间质干细胞(MSCs)对CD4(+)CD25(+)Treg细胞的影响。方法:采用Ficoll分离ITP患者及健康对照者骨髓单个核细胞,通过体外培养,扩增出MSCs。通过Ficoll分离法和尼龙棉柱法获取ITP患者外周血T淋巴细胞,ITP患者MSCs经丝裂霉素C处理后按不同数量(1×103、1×104、1×105个细胞/孔)(A1、A2、A3组)接种培养板作为基底层细胞;健康对照者MSCs经丝裂霉素C处理后按不同数量(1×103、1×104、1×105个细胞/孔)(B1、B2、B3组)接种培养板作为基底层细胞;然后分别以2×105个细胞/孔接种体外分离纯化的ITP患者T淋巴细胞,设立空白组C组(ITP患者T淋巴细胞单独培养)。培养第3天各自收集T淋巴细胞,用流式细胞术检测各组CD4(+)CD25(+)Treg细胞比例。结果:在血凝素作用下,实验组及对照组各浓度亚组CD4(+)CD25(+)Treg细胞数量均较空白组明显减少(P<0.05);A2组较B2组CD4(+)CD25(+)Treg细胞数量显著减少(P<0.05),A3组较A2、B3组CD4(+)CD25(+)Treg细胞数量明显减少(P<0.05)。结论:ITP患者可能存在MSCs质或量的改变,导致免疫调节功能异常,对CD4(+)CD25(+)Treg细胞增殖抑制作用明显加强,CD4(+)CD25(+)Treg细胞的不足引起外周免疫耐受减弱,MSCs的免疫调节功能异常可能在ITP发病过程中起重要作用。Objective：To investigate the effect of mesenchymal stem cells（MSCs）on CD4（＋）CD25（＋）Treg cells proliferation in patients with ITP in vitro.Method：Bone marrow from ITP patients and healthy persons derived MSCs were isolated by Ficoll Hypaque and cultured for proliferating to passage cells.T lymphocytes of ITP patients were isolated from peripheral blood by Ficoll Hypaque and nylon cotton column.Then the stromal feeder layers of different numbers（1×10^3,1×10^4,1×10^5 per well）of MSCs treated with mitomycin were co-cultured with the T lymphocytes（2×10^5 per well）.The proportions of CD4（＋）CD25（＋）Treg cells controls were determined by flow cytometry after 3days cultured.Result：The proportions of CD4（＋）CD25（＋）Treg cells in experimental group and control group were all lower than that in non-treatment control group.The proportions of CD4（＋）CD25（＋）Treg cells in experimental group were lower than those in control group when MSCs number was1×10^4,1×10^5 per well.Conclusion：CD4（＋）CD25（＋）Treg cells may play an important role in the pathogenesis of ITP.The quality or quantity of MSCs in ITP patients may alter,and this change may lead to immune dysfunction.

关 键 词：骨髓间质干细胞 免疫性血小板减少症 CD4(+)CD25(+)Treg细胞 

分 类 号：R558.2[医药卫生—血液循环系统疾病]