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/mL,PBMC group was (0.90±0.05) pg/mL,PBMC+MSC group was (11.00±3.02) pg/mL and CD3+CD28+PBMC group was (6.04±1.90) pg/mL,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 +CD45RA-CD49b+LAG3+Tr1 in PBMC which stimulated by CD3 and CD28.The increase of IL-10 was related to the increase of CD4+CD45RA-C D49b+LAG3 +Tr 1 proportion.%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+CD45RA-CD49b+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+C D45 RA-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/mL,PBMC组为(0.90±0.05)pg/mL,PBMC+MSC组为(11.00±3.02)pg/mL,CD3+CD28+PBMC组为(6.04± 1.90)pg/mL,差异有统计学意义(P＜0.05).MSC+CD3+CD28+PBMC组中的CD4+CD45RA-CD49b+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.
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