摘要
目的探讨用钙离子载体A23187通过钙动员机制使人急性粒细胞性白血病细胞系HL-60细胞迅速分化为树突状细胞（DCs）的方法，为今后临床用DCs进行抗白血病免疫治疗打下基础。方法将生长良好的HL-60细胞加入不同浓度A23187（45～720ng／mL）或联用重组人γ-干扰素（rhIFN-γ，1000U／mL）培养20～96h，倒置显微镜和扫描电镜观察其形态学特征，流式细胞术检测其免疫表型，混合淋巴细胞反应检测其刺激T淋巴细胞增殖的能力。结果HL-60细胞加入适量A23187（180ng／mL）或联合rhIFN-γ（1000 U／mL）诱导20h后，即可出现细胞表面树状突起，且树突状细胞表面特异性成熟标志抗原CD83、共刺激分子CD80、CD86表达升高；培养48h，CD83分子表达开始降低；培养72h，可获得DC的典型形态，CD80、CD86分子表达持续升高。同种异体混合淋巴细胞反应检测经A23187或联用rhIFN-γ诱生的DCs可明显刺激同种异体T淋巴细胞增殖。结论钙离子载体可诱导HL-60细胞分化为高表达共刺激分子及具有刺激同种异体T淋巴细胞增殖的成熟DCs，钙动员有望成为一种快速获取DCs的有效方法。
Objective To investigate the method for calcium mobilization inducing rapidly to produce the dendritic cells （DC） from myeloid leukemia cells. Methods HL-60 cells were cultured with calcium ionophore A23187 （45-720 ng/mL） or plus rhIYN-γ （1000 U/mL） for 20-96 hours. The morphologie features of cells were observed under inverted microscope and scanning electron microscope, when the cell phenotypes of HL-60 treated with A23187 were determined by flow eytometry. The proliferation of allogeneie human T cells was tested by mixed lymphocyte reaction （Allo-MLR）. Results After treated with A23187 （180 ng/mL） or plus rhIFN-γ （1000 U/mL） for 20 hours, some of HL-60 cells were found to have the dendritic appearance on cell surfaee; and the CD83 as a characteristic marker of mature DCs, co-stimulating molecules CD80 and CD86 got an up-regulated expression. When HL-60 cells were cultured with A23187 for 48 hours, the expression of CD83 began going down. A large number of cells with typical dendritic appearance were observed after cultured with A23187 for 72 hours, and the expressions of CD80 and CD86 were up-regulated continuously. Allo-MLR revealed that DCs derived from HL-60 cells treated with A23187 or plus rhIFN-γ could stimulate the proliferation of allogeneic human T cells. Conclusion The calcium ionophore A23187 can induce the HL-60 cells into mature DCs. This study suggests that calcium mobilization for cell differentiation may be an alternative way to rapidly acquire DCs.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2007年第2期209-212,共4页
Journal of Sichuan University(Medical Sciences)
基金
四川省科技厅基金(批准号02SY029-143)
四川省学术带头人培养基金(批准号06SXS-10)资助
