【摘要】 目的研究人胎盘来源的间充质干细胞(MSC)是否能诱导正常人外周血单个核细胞(PBMC)中CD4+CD45RACD49b+LAG3+Tr1细胞增殖。方法分离、纯化、表型鉴定人胎盘来源的MSC。实验分组如下:PBMC组、PBMC+MSC组、CD3+CD28+PBMC组和MSC+CD3+CD28+PBMC组。相同条件下培养24 h后,流式细胞术检测各组CD4~+CD45RA~-CD49b~+LAG3~+Tr1细胞比例;ELISA法检测各组上清中白细胞介素10(IL-10)水平,并对MSC+CD3+CD28+PBMC组中的CD4~+CD45RA~-CD49b~+LAG3~+Tr1比例和其上清中IL-10含量作相关性分析。结果MSC+CD3+CD28+PBMC组中CD4~+CD45RA~-CD49b~+LAG3~+Tr1细胞比例为(6.66±2.80)%,PBMC组为(1.01±0.44)%,PBMC+MSC组为(1.14±0.78)%,CD3+CD28+PBMC组为(4.20±2.58)%,差异有统计学意义(P<0.05)。MSC+CD3+CD28+PBMC组上清中的IL-10含量为(14.45±3.14)pg/m L,PBMC组为(0.90±0.05)pg/m L,PBMC+MSC组为(11.00±3.02)pg/m L,CD3+CD28+PBMC组为(6.04±1.90)pg/m L,差异有统计学意义(P<0.05)。MSC+CD3+CD28+PBMC组中的CD4+CD45RACD49b+LAG3+Tr1比例与其上清中IL-10含量成正相关(r=0.880,P<0.01)。结论 MSC可以诱导CD3、CD28活化的PBMC细胞中的CD4~+CD45RA~-CD49b~+LAG3~+Tr1细胞增殖,CD4~+CD45RA~-CD49b~+LAG3~+Tr1细胞比例增加与上清中IL-10水平升高有关。更多还原

【Abstract】 Objective To investigate the proliferative effect of human-placenta derived mesenchymal stem cells(MSC)on the CD4~+CD45RA~-CD49b~+LAG3~+Tr1 in normal human peripheral blood cells(PBMC). Methods MSCs were isolated from placental tissue, enriched and expandedby using adherent culture. Four different groups were set up: PBMC group,PBMC +MSC group, CD3 +CD28 +PBMC group, and MSC +CD3 +CD28 +PBMC group. 24 hours after training under the same condition, the proportion of CD4~+CD45RA~-CD49b~+LAG3~+Tr1 was detected by flow cytometry and the levels of interleukin 10(IL-10) were detected by ELISA. Results The proportion of CD4~+CD45RA~-CD49b~+LAG3~+Tr1 of MSC +CD3+CD28+PBMC group was(6.66±2.80)%, the proportion of PBMC group was(1.01±0.44)%, the proportion of PBMC+MSC group was(1.14±0.78)%, and the proportion of CD3+CD28+PBMC group was(4.20±2.58)%, with statistically significant difference(P < 0.05). The IL-10 level of MSC+CD3+CD28+PBMC group was(14.45±3.14) pg/m L, PBMC group was(0.90±0.05) pg/m L, PBMC+MSC group was(11.00±3.02) pg/m L and CD3+CD28+PBMC group was(6.04±1.90) pg/m L,with statistically significant difference(P < 0.05). The level of IL-10 had a significant lypositive correlation with the proportion of CD4~+CD45RA~-CD49b~+LAG3~+Tr1 in MSC +CD3 +CD28 +PBMC group(r =0.880, P < 0.01). Conclusion MSC can promote the proliferation of CD4+CD45RACD49b+LAG3+Tr1 in PBMC which stimulated by CD3 and CD28. The increase of IL-10 was related to the increase of CD4~+CD45RA~-CD49b~+LAG3~+Tr1proportion.更多还原