钙离子载体促进K562细胞诱导分化成DC样细胞的研究

Differentiation of K562 cells into DC-like cells induced by the combination of calcium ionorphore with rhGM-CSF and rhIL-4

目的观察钙离子载体(CI)诱导慢性髓系白血病细胞株K562分化成DC样细胞的作用。方法将细胞分三组培养:第1组加入GM-CSF1000U/ml和IL-4500U/ml;第2组加入GM-CSF1000U/ml、IL-4500U/ml和CI375ng/ml;第3组为对照,不加细胞因子,也不加CI。流式细胞仪检测各组细胞表面免疫表型的表达,MTT比色法检测其刺激同种异体T细胞增殖的作用。结果细胞因子加CI组培养48h后即可见细胞形态呈多形性,有些细胞可见突起;96h后CD80、CD86、CD83的表达率较对照组、单用细胞因子明显提高,明显刺激同种异体T淋巴细胞增殖。单用细胞因子组培养96h后细胞形态无明显变化,CD80、CD86的阳性率明显高于对照组,CD83与对照组之间差异无统计学意义,不能明显刺激同种异体T淋巴细胞增殖。结论CI与GM-CSF和IL-4联合可迅速将K562细胞诱导成DC样细胞;CI与细胞因子可能有协同作用。

Objective To investigate the possibility of calcium ionorphore （CI） in inducing K562 cell differentiation into dendritic cell （DC） -like cells. Methods K562 cells in bloom were cultured in the medium with cytokines （ GM-CSF and IL-4） （ group 1 ）, with same cytokines added and CI （ group 2） and control medium （ without cytokines and CI, group 3 ）, respectively. The terminal concentration of GM-CSF, IL- 4 and CI were 1 000 U/ml, 500 U/ml, 375 ng/ml, respectively. Surface immunologic phenotypes were analyzed by flow cytometry and the proliferation of allogeneic T cells by MTT colorimetry. Results In group 2, polymorphologic changes of K562 cells, including the formation of processes ,were observed at 48 h. CD80, CD86 and CD83 were more markedly up-regulated than group 3 and group 1. The effect of group 2 cells in stimulating allogeneic T cells was more significant than the other two groups. In group 1, no notable morphological change was found at 96 h of cell culture. CD80, CD86 were more markedly up-regulated than group 3. There was no notable difference in the expression rate of CD83 in group 3 and group 1. No marked increase of human T cells proliferation was detected compared with group 3. Conclusions Combination of CI with GM-CSF and IL-4 can rapidly induce the K562 cells to differentiate into DC-like cells. These cytokines and CI might have synergistic effect.