基础医学与临床 ›› 2011, Vol. 31 ›› Issue (10): 1134-1138.

• 研究论文 • 上一篇    下一篇

脐血来源间充质干细胞可抑制异体T细胞的反应究

李志斌   

  1. 陕西省妇幼保健院
  • 收稿日期:2010-08-23 修回日期:2010-12-22 出版日期:2011-10-05 发布日期:2011-10-08
  • 通讯作者: 李志斌 E-mail:lzhbzyzh@163.com

The inhibitory effects of human umbilical cord blood derived mesenchymal stem cells on allogeneic peripheral blood T lymphocytes

  • Received:2010-08-23 Revised:2010-12-22 Online:2011-10-05 Published:2011-10-08

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CD3+T与HUCB-MSCs 1:1混合培养5 天,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,表达CD29、CD44、HLA-ABC。HUCB-MSCs不刺激T细胞增殖 (3760 ± 730 counts/min versus 3600 ± 800 counts/min,p=0.85),但抑制PHA引起的T细胞增殖 (5230 ± 550 counts/min versus 10500 ± 800 counts/min,p<0.001);HUCB-MSCs能抑制异体T细胞分泌IFN-γ(510 ± 60 pg/ml versus 1580 ± 100 pg/ml, p<0.001)和 TNF-α(590 ± 20 pg/ml versus 1180 ± 30 pg/ml, p<0.001),上调IL-4 (16.3 ± 8.2 pg/ml versus 4.1 ± 1.8 pg/ml, p<0.001) 和 IL-10(105 ± 5 pg/ml versus 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。

关键词: 脐血, 间充质干细胞, T淋巴细胞, 抑制

Abstract: Objective To study the influences of human umbilical cord blood derived mesenchymal stem cells (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 were determinate 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 were investigated 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 failed to elicit positive responses of T cells (3760 ± 730 counts/min versus 3600 ± 800 counts/min,p=0.85), whereas significantly suppressed PHA-stimulated T-cell proliferation(5230 ± 550 counts/min versus 10500 ± 800 counts/min,p<0.001). Both pro-inflammatory cytokines IFN-γ(510 ± 60 pg/ml versus 1580 ± 100 pg/ml, p<0.001)and TNF-α(590 ± 20 pg/ml versus 1180 ± 30 pg/ml, p<0.001)secreted by T cells were inhibited by HUCB-MSCs while anti-inflammatory cytokines IL-10(105 ± 5 pg/ml versus 17 ± 2 pg/ml, p<0.001)and IL-4 (16.3 ± 8.2 pg/ml versus 4.1 ± 1.8 pg/ml, p<0.001) were up-regulated. In addition, MSCs-inhibited T cells were not apoptotic during 5 days’ culture. Conclusion HUCB-MSCs possessed could inhibit the activity of allogeneic peripheral blood T lymphocytes.

Key words: umbilical cord blood, mesenchymal stem cells, T lymphocytes, inhibition

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