miRNA-155对调节性T细胞表型和功能的影响
发布时间：2018-10-16 09:38
【摘要】：目的探讨微小核糖核酸(miRNA)-155对调节性T细胞(Treg)的两种亚型诱导性Treg(iTreg)和天然性Treg(nTreg)的影响。方法采用健康成人外周血分离获取的外周血单个核细胞(PBMC),利用磁性细胞分选法分别获取幼稚T细胞和nTreg。培养阶段将细胞分为3组:对照组(幼稚T细胞加入白细胞介素-2培养)、iTreg组(幼稚T细胞加入白细胞介素-2和转化生长因子-β培养)和nTreg组(nTreg加入白细胞介素-2培养)。每组再分为3个亚组:未处理亚组、scramble亚组和miRNA-155拮抗剂亚组(每亚组3个孔)。采用低密度芯片分析方法检测3组中的未处理亚组细胞中miRNA-155的基因表达水平。采用流式细胞术检测3组中各亚组细胞的表面标志物CD25、Foxp3、CD127水平。采用流式细胞术检测3组中各亚组细胞的CD4+CD25+Foxp3+SOCS1+Treg比例;采用流式细胞术检测3组中各亚组细胞的Treg抑制功能。结果与对照组和iTreg组比较,nTreg组细胞的miRNA-155表达水平明显降低,差异有统计学意义(均为P0.05)。与对照组和iTreg组比较,nTreg组的SOCS1表达水平明显升高,差异均有统计学意义(均为P0.05)。加入miRNA-155拮抗剂后并未导致Foxp3、CD127和CD25等Treg重要表面标志物发生明显的变化。与对照组和iTreg组比较,nTreg组的SOCS1表达水平明显升高,差异均有统计学意义(均为P0.05)。iTreg组中的未处理亚组细胞miRNA-155表达水平较低,而拮抗剂抑制其表达之后(miRNA-155拮抗剂亚组),能够使其SOCS1表达升高。iTreg组中,与未处理亚组比较,miRNA-155拮抗剂亚组的Treg抑制功能在1∶8、1∶16、1∶32的比例时,显示出了更强的抑制功能(均为P0.05)。结论体外拮抗miRNA-155对nTreg的抑制功能无明显影响,但是能够增加iTreg的SOCS1表达水平和体外抑制功能。
[Abstract]:Objective to investigate the effects of (miRNA) 155 on two subtypes of regulatory T cell (Treg), inducible Treg (iTreg) and natural Treg (nTreg). Methods (PBMC), of peripheral blood mononuclear cells isolated from healthy adults was used to obtain immature T cells and nTreg. by magnetic cell sorting method. At the culture stage, the cells were divided into three groups: control group (immature T cells cultured with interleukin-2 (), iTreg) group (immature T cells added interleukin-2 and transforming growth factor- 尾 culture) and nTreg group (nTreg supplemented with IL-2 culture). Each group was subdivided into 3 subgroups: untreated subgroup, scramble subgroup and miRNA-155 antagonist subgroup (3 holes per subgroup). Low density microarray analysis was used to detect the expression of miRNA-155 gene in untreated subgroups of three groups. Flow cytometry was used to detect the level of surface marker CD25,Foxp3,CD127 in each subgroup of the three groups. Flow cytometry was used to detect the proportion of CD4 CD25 Foxp3 SOCS1 Treg in each subgroup of the three groups, and flow cytometry was used to detect the inhibitory function of Treg in each subgroup of the three groups. Results compared with control group and iTreg group, the expression of miRNA-155 in nTreg group was significantly lower than that in control group and iTreg group (P0.05). Compared with control group and iTreg group, the expression of SOCS1 in nTreg group was significantly higher than that in control group and iTreg group (P0.05). The addition of miRNA-155 antagonist did not cause significant changes in Treg surface markers such as Foxp3,CD127 and CD25. Compared with the control group and the iTreg group, the expression of SOCS1 in the nTreg group was significantly higher than that in the control group and iTreg group, and the difference was statistically significant (P0.05) the expression of miRNA-155 in the untreated subgroup was lower than that in the). ITreg group. After the antagonist inhibited its expression (miRNA-155 antagonist subgroup), the SOCS1 expression was increased. In iTreg group, compared with untreated subgroup, the Treg inhibitory function of miRNA-155 antagonist subgroup was 1: 8: 1: 161: 32. It showed stronger inhibitory function (P0.05). Conclusion antagonizing miRNA-155 in vitro has no obvious effect on the inhibitory function of nTreg, but it can increase the SOCS1 expression level of iTreg and its inhibitory function in vitro.
【作者单位】： 南京医科大学第一附属医院肝脏外科;
【基金】：国家自然科学基金(81273262、81210108017、81100270、81070380)
【分类号】：R392
本文编号：2273950
[Abstract]:Objective to investigate the effects of (miRNA) 155 on two subtypes of regulatory T cell (Treg), inducible Treg (iTreg) and natural Treg (nTreg). Methods (PBMC), of peripheral blood mononuclear cells isolated from healthy adults was used to obtain immature T cells and nTreg. by magnetic cell sorting method. At the culture stage, the cells were divided into three groups: control group (immature T cells cultured with interleukin-2 (), iTreg) group (immature T cells added interleukin-2 and transforming growth factor- 尾 culture) and nTreg group (nTreg supplemented with IL-2 culture). Each group was subdivided into 3 subgroups: untreated subgroup, scramble subgroup and miRNA-155 antagonist subgroup (3 holes per subgroup). Low density microarray analysis was used to detect the expression of miRNA-155 gene in untreated subgroups of three groups. Flow cytometry was used to detect the level of surface marker CD25,Foxp3,CD127 in each subgroup of the three groups. Flow cytometry was used to detect the proportion of CD4 CD25 Foxp3 SOCS1 Treg in each subgroup of the three groups, and flow cytometry was used to detect the inhibitory function of Treg in each subgroup of the three groups. Results compared with control group and iTreg group, the expression of miRNA-155 in nTreg group was significantly lower than that in control group and iTreg group (P0.05). Compared with control group and iTreg group, the expression of SOCS1 in nTreg group was significantly higher than that in control group and iTreg group (P0.05). The addition of miRNA-155 antagonist did not cause significant changes in Treg surface markers such as Foxp3,CD127 and CD25. Compared with the control group and the iTreg group, the expression of SOCS1 in the nTreg group was significantly higher than that in the control group and iTreg group, and the difference was statistically significant (P0.05) the expression of miRNA-155 in the untreated subgroup was lower than that in the). ITreg group. After the antagonist inhibited its expression (miRNA-155 antagonist subgroup), the SOCS1 expression was increased. In iTreg group, compared with untreated subgroup, the Treg inhibitory function of miRNA-155 antagonist subgroup was 1: 8: 1: 161: 32. It showed stronger inhibitory function (P0.05). Conclusion antagonizing miRNA-155 in vitro has no obvious effect on the inhibitory function of nTreg, but it can increase the SOCS1 expression level of iTreg and its inhibitory function in vitro.
【作者单位】： 南京医科大学第一附属医院肝脏外科;
【基金】：国家自然科学基金(81273262、81210108017、81100270、81070380)
【分类号】：R392
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