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目的分析胞外泛素对Treg细胞发挥免疫抑制作用的影响及其可能机制,初步分析临床病人输注血制品中泛素浓度与其TRIM发生的相关性。方法用美天旎磁珠试剂盒分离CD4+CD25+CD127-Treg细胞,与FITC荧光标记泛素共孵育(37℃孵育箱,2 h),流式细胞仪分析泛素与Treg的结合;Transwell分析泛素对人Treg细胞的趋化迁移的影响;美天旎磁珠分选CD4+CD25+Foxp3+Treg细胞、效应性T细胞按1∶1共培养,加入CD3+CD28beads刺激Treg细
Objective To analyze the effect of extracellular ubiquitin on immunosuppressive effect of Treg cells and its possible mechanism, and to analyze the correlation between the concentration of ubiquitin and the incidence of TRIM in clinical patients. Methods The CD4 + CD25 + CD127-Treg cells were isolated by Metitian magnetic beads kit and incubated with FITC fluorescent labeled ubiquitin (37 ℃ incubator for 2 h). The binding of ubiquitin to Treg was analyzed by flow cytometry. Transwell The effect of ubiquitin on chemotactic migration of human Treg cells was analyzed. Meantime magnetic beads were used to sort CD4 + CD25 + Foxp3 + Treg cells. Effector T cells were co-cultured at 1: 1, and CD3 + CD28beads were added to stimulate Treg cells