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目的:观察白细胞介素21(interleukin-21,IL-21)对慢性粒细胞白血病(chronic myeloid leukemia,CML)患者CD4+CD25+调节性T细胞(regulatory T cell,Treg)的作用,为CML的免疫治疗提供新思路。方法:收集兰州大学第二医院血液科初诊CML患者外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),免疫磁珠法分选出CD4+T细胞。分别加入50、100、200 ng/ml IL-21与CD4+T细胞共培养72 h后,流式细胞术检测Treg的比例,并设生理盐水空白对照组。Real-time PCR法检测Treg细胞中Foxp3 mRNA的表达,ELISA法检测各组细胞上清液中IL-10、TGF-β的水平。结果:与空白对照组相比,50、100、200 ng/ml IL-21组CD4+CD25+Treg占CD4+T细胞的比例均降低[(3.42±0.76)%、(6.81±0.33)%、(7.98±0.76)%vs(12.09±0.91)%,P<0.05];且3组的Foxp3 mRNA均降低[(0.05±0.02)、(0.16±0.02)、(0.25±0.02)vs 1,P<0.05];Treg分泌IL-10量减少[(26.13±7.28)、(44.88±3.72)、(79.77±3.94)vs(133.00±12.32)pg/ml,P<0.05],分泌TGF-β量也减少[(9.25±0.84)、(16.70±1.00)、(20.47±1.60)vs(26.05±1.81)pg/ml,P<0.05]。结论:IL-21在体外可减少CML患者外周血中Treg的比例,并抑制其功能。
Objective: To observe the effect of interleukin-21 (IL-21) on CD4 + CD25 + regulatory T cell (Treg) in patients with chronic myeloid leukemia (CML) Treatment provides new ideas. Methods: Peripheral blood mononuclear cells (PBMCs) from newly diagnosed CML patients were collected from Department of Hematology, Second Hospital of Lanzhou University. CD4 + T cells were sorted by immunomagnetic beads method. After co-cultured with 50, 100 and 200 ng / ml IL-21 and CD4 + T cells for 72 h respectively, the proportion of Tregs was detected by flow cytometry and normal saline control group. The expression of Foxp3 mRNA in Treg cells was detected by Real-time PCR. The levels of IL-10 and TGF-β in the supernatant of each group were detected by ELISA. Results: Compared with the blank control group, the proportion of CD4 + CD25 + Tregs in CD4 + T cells in 50,100,200 ng / ml IL-21 group was significantly lower than that in control group [(3.42 ± 0.76)%, (6.81 ± 0.33)%, (7.98 ± 0.76)% vs (12.09 ± 0.91)% respectively, P <0.05]. The Foxp3 mRNA in the three groups were significantly lower than that in the control group [(0.05 ± 0.02) vs (0.16 ± 0.02) vs 0.05]. The amount of IL-10 secreted by Treg decreased (26.13 ± 7.28), (44.88 ± 3.72), (79.77 ± 3.94) vs (133.00 ± 12.32) pg / ml, [(9.25 ± 0.84), (16.70 ± 1.00), (20.47 ± 1.60) vs (26.05 ± 1.81) pg / ml, P <0.05]. Conclusion: IL-21 can reduce the proportion of Treg in peripheral blood of CML patients in vitro and inhibit its function.