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本研究旨在探索钙载体(calcium ionophore, CI)诱导慢性髓性白血病(CML)细胞分化成树突状细胞(DC)的作用,了解诱导产生P210蛋白的情况和刺激自身T细胞产生针对CML细胞的细胞毒作用。用CI和粒-单集落刺激因子(GM-CSF)联合诱导CML病人骨髓来源的单个核细胞,通过倒置显微镜和电子显微镜观察细胞的形态变化和超微结构,用流式细胞仪检测细胞表面分化抗原的变化和变化过程;用Western blot检测CML特异的肿瘤标志物P210蛋白在DC中的表达;通过乳酸脱氢酶(LDH)实验评估CI诱导的CML-DC刺激自身T细胞产生抗CML细胞的细胞毒作用。结果表明: CML细胞经CI和GM-CSF联合诱导96小时后在倒置显微镜和电子显微镜下观察到DC典型的形态结构;流式细胞仪检测显示CD83、CD86、CD80、HLA-DR、CD40等细胞表面分化抗原的表达显著提高;Western blot实验表明,生成的CML-DC有P210蛋白的表达,与未经诱导的CML单个核细胞比较,用CI和GM-CSF联合诱导生成的CML-DC的P210蛋白表达水平降低;LDH实验表明,这些CML-DC刺激的自身T细胞对CML细胞的杀伤率显著高于用CML细胞刺激的自身T细胞。结论: CI和GM-CSF联合能快速诱导CML细胞分化成DC,这些CML-DC表达CML细胞特异的肿瘤标志物P210蛋白,但其表达水平较未经诱导的CML单个核细胞低;CML细胞经诱导生成的CML-DC能够刺激自身T细胞产生抗白血病细胞毒性。
The purpose of this study was to explore the role of calcium ionophore (CI) in the differentiation of chronic myeloid leukemia (CML) cells into dendritic cells (DCs), to understand the induction of P210 protein production and to stimulate self T cell production against CML cells Cytotoxicity. Bone marrow-derived mononuclear cells from CML patients were induced by combination of CI and granulocyte-single colony stimulating factor (GM-CSF). The morphological changes and ultrastructure of the cells were observed by inverted microscope and electron microscope. Cell surface differentiation was detected by flow cytometry The changes and changes of antigens were detected by Western blot. The expression of CML-specific tumor marker P210 in DCs was detected by Western blot. CI-induced CML-DCs stimulated autologous T cells to produce anti-CML cells by lactate dehydrogenase (LDH) assay Cytotoxic effects. The results showed that typical morphological structures of DCs were observed under inverted microscope and electron microscope after CML cells were induced by CI and GM-CSF for 96 hours. Flow cytometry showed that CD83, CD86, CD80, HLA-DR, CD40 and other cells Western blot analysis showed that P210 protein was expressed in CML-DCs. Compared with non-induced CML mononuclear cells, P210 of CML-DCs induced by CI and GM-CSF were significantly increased LDH test showed that the killing rate of CML-DCs stimulated by these CML-DCs on CML cells was significantly higher than that of CML-stimulated T cells. Conclusions: The combination of CI and GM-CSF can rapidly induce the differentiation of CML cells into DC. These CML-DCs express CML cell-specific tumor marker P210 protein, but their expression level is lower than that of non-induced CML mononuclear cells. CML cells The induced CML-DC can stimulate its own T cells to produce anti-leukemia cytotoxicity.