摘要
目的研究脐血间充质干细胞(HUCB-MSCs)对异体T细胞的抑制作用。方法体外培养HUCB-MSCs,流式细胞术测表面标记;取正常人外周血,免疫磁珠分离CD3+T细胞,将分离的CD3+T与HUCB-MSCs 1∶1混合培养5 d,PHA刺激或不刺激,采用3H-TdR掺入法观察T细胞增殖,ELISA方法检测细胞因子,流式细胞术观察细胞凋亡。结果 HUCB-MSCs呈纺锤样的细胞形态,不表达CD14、CD34、CD45、HLA-DR,而表达CD29、CD44、HLA-ABC。HUCB-MSCs抑制PHA引起的T细胞增殖(5 230±550 vs 10 500±800 counts/min,P<0.001);HUCB-MSCs还能抑制异体T细胞分泌IFN-γ(510±60 vs 1 580±100 pg/mL,P<0.001)和TNF-α(590±20 vs 1 180±30 pg/mL,P<0.001),上调IL-4(16.3±8.2 vs 4.1±1.8 pg/mL,P<0.001)和IL-10(105±5 vs 17±2 pg/mL,P<0.001)分泌;HUCB-MSCs不诱导T细胞的凋亡。结论 HUCB-MSCs能抑制异体T细胞免疫反应。
Objective To study the influences of mesenchymal stem cells derived from human umbilical cord blood（HUCB-MSCs） on allogeneic peripheral blood T lymphocytes.Methods HUCB-MSCs were isolated from normal human umbilical cord blood,isolated and cultured in vitro,and then surface marker was examined with FCM.After isolating CD3＋T cells from normal human peripheral blood,CD3＋T cells were mixed with HUCB-MSCs stimulated with PHA or not.Five days later,the effects of HUCB-MSCs on T-cell proliferation was detected by 3H-TdR.Cytokines production of PHA-stimulated T cells in co-culture was measured with ELISA.T-cell apoptosis affected by HUCB-MSCs were also observed by FCM.Results HUCB-MSCs showed a spindle-shaped fibroblastic morphology in culture.The populations appeared as a homogenous population and were uniformly positive for the expression of CD29,CD44,and HLA-ABC,but negative for the expression of CD14,CD34,CD45,and HLA-DR.In co-culture,MSCs significantly suppressed PHA-stimulated T-cell proliferation（5 230±550 vs 10 500±800 counts/min,P0.001）.Both pro-inflammatory cytokines IFN-γ（510±60 vs 1 580±100 pg/mL,P0.001）and TNF-α（590±20 vs 1 180±30 pg/mL,P0.001）secreted by T cells were inhibited by HUCB-MSCs while anti-inflammatory cytokines IL-10（105±5 vs 17±2 pg/mL,P0.001）and IL-4（16.3±8.2 vs 4.1±1.8 pg/mL,P0.001） were up-regulated.In addition,MSCs-inhibited T cells were not apoptotic during 5 days＇ culture.Conclusions HUCB-MSCs may inhibit the activity of allogeneic peripheral blood T lymphocytes.
出处
《基础医学与临床》
CSCD
北大核心
2011年第10期1134-1138,共5页
Basic and Clinical Medicine
关键词
脐血
间充质干细胞
T淋巴细胞
抑制
umbilical cord blood
mesenchymal stem cells
T lymphocytes
inhibition
