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目的探讨重组高迁移率族蛋白1(r HMGB1)对髓源性抑制性细胞(MDSC)的体外分化作用。方法分离BALB/c小鼠的骨髓细胞,分别使用r HMGB1、粒细胞-巨噬细胞集落刺激因子联合白细胞介素6(GM-CSF-IL-6)、GM-CSF、IL-6和r HMGB1三者联合(GM-CSF-IL-6-r HMGB1)进行体外刺激48 h,流式细胞术检测CD11b+Gr1+MDSC、CD11c+和F4/80+巨噬细胞的比例。用免疫磁珠体外分选出粒细胞源性MDSC(G-MDSC)和巨噬细胞源性MDSC(M-MDSC)分别用r HMGB1刺激48 h,流式细胞术检测各组细胞中CD11b+Gr1+MDSC、CD11c+细胞和F4/80+巨噬细胞的比例。结果与对照组相比,r HMGB1、GM-CSF-IL-6、GM-CSF-IL-6-HMGB1刺激48 h后,CD11b+Gr1+MDSC比例增加,CD11c+细胞和F4/80+巨噬细胞的比例减少;免疫磁珠体外分选出G-MDSC和M-MDSC,用r HMGB1蛋白刺激48 h,与对照组相比,r HMGB1刺激后,CD11b+Gr1+MDSC、CD11c+细胞和F4/80+巨噬细胞的比例无显著性差异。结论 r HMGB1体外可以诱导分化MDSC比例增加,GM-CSF、IL-6与r HMGB1联用可以增强诱导效果。
Objective To investigate the effect of r HMGB1 on the differentiation of myeloid derived inhibitory cells (MDSCs) in vitro. Methods The myeloid cells of BALB / c mice were isolated and treated with HMGB1, GM-CSF-IL-6, GM-CSF, IL-6 and HMGB1 GM-CSF-IL-6-r HMGB1 was stimulated in vitro for 48 h. The proportion of CD11b + Gr1 + MDSC, CD11c + and F4 / 80 + macrophages was detected by flow cytometry. The granulocyte-derived MDSC (G-MDSC) and macrophage-derived MDSC (M-MDSC) were sorted by immunomagnetic beads in vitro for 48 h after stimulation with r HMGB1. Flow cytometry was used to detect the expression of CD11b + Gr1 + MDSC, CD11c + cells and F4 / 80 + macrophages. Results Compared with the control group, the proportion of CD11b + Gr1 + MDSC increased after 48h stimulation with r HMGB1, GM-CSF-IL-6 and GM-CSF-IL-6-HMGB1, The percentage of CD11b + Gr1 + MDSC, CD11c + cells and F4 / 80 cells were significantly decreased after r HMGB1 stimulation compared with the control group. Immunomagnetic beads were used to separate G-MDSC and M-MDSC in vitro and stimulated with r HMGB1 for 48 h. + Macrophages ratio was no significant difference. Conclusions r HMGB1 can induce the differentiation of MDSC in vitro, and GM-CSF, IL-6 and r HMGB1 can enhance the induction effect.