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本研究观察低温保存对白血病源性树突状细胞(L-DCs)的生物学特性和功能的影响。取急性、慢性髓系白血病骨髓细胞,将其一部分细胞立即检测,一部分细胞立即培养,一部分细胞加入细胞保护剂低温保存一定时间复温后培养。细胞培养时,在细胞培养体系内加入组合细胞因子并培养12天,然后分别检测3组细胞的细胞形态、细胞免疫表型、混合淋巴细胞反应及细胞毒T淋巴细胞(CTL)效应,并对结果进行相互比较分析。结果表明:在细胞形态学方面,急性、慢性髓系白血病患者骨髓细胞在不染色或在瑞氏染色下仅见白血病细胞,而未观察到“树突状”细胞,但经组合的细胞因子培养后,均出现了“树突状”形态的细胞,低温保存前、后二者比较,无明显差异;在细胞表面免疫标志表达方面,经组合细胞因子培养的细胞与未培养的细胞相比,细胞表面表达的CD80、CD54、HLA-DR、CD1a、CD83、CD86明显上调,而CD14表达则明显下调,低温保存前、后的细胞在上述几种细胞表面免疫表达指标方面相比较无明显差异;低温保存后的白血病细胞经培养后,不仅有明显的混合淋巴细胞反应,而且在与T细胞作用后T细胞表面抗原CD8和CD25免疫标记细胞明显增多,明显地表现了CTL杀伤自体白血病细胞的效应。结论:髓系白血病细胞在组合细胞因子培养条件下,可诱生为L-DC;L-DC的诱生在生物学特性方面不受低温保存的影响。
This study was to observe the effects of cryopreservation on the biological characteristics and function of leukemic DCs. Acute, chronic myeloid leukemia bone marrow cells, some of its cells immediately detected, some cells immediately cultured, some of the cells added cytoprotective agent cryogenic preservation of a certain time after rewarming culture. At the time of cell culture, the combined cytokines were added into the cell culture system and cultured for 12 days. Then the cell morphology, cellular phenotype, mixed lymphocyte reaction and cytotoxic T lymphocyte (CTL) The results were compared with each other. The results showed that, in terms of cell morphology, bone marrow cells from patients with acute, chronic myeloid leukemia either did not stain or showed only leukemic cells under Wright’s staining, while no “dendritic” cells were observed, but combined cytokines There were no significant differences between the two groups in terms of “dendritic” morphology after culture, and no significant difference was observed between the two groups before and after cryopreservation; in terms of the expression of cell surface immunological markers, the cells cultured with the combination of cytokines and the cells without culture CD80, CD54, HLA-DR, CD1a, CD83 and CD86 on the cell surface were significantly up-regulated while the expression of CD14 was significantly down-regulated compared with that on the cell surface before and after cryopreservation There was a significant difference between the two groups. After leukemia cells were cultured, not only the mixed lymphocyte reaction but also the number of CD8 and CD25 immunostaining cells significantly increased after treated with T cells. CTL anti-autologous leukemia The effect of cells. CONCLUSION: Myeloid leukemia cells can be induced to L-DC under the combined cytokine culture conditions. L-DC induction is not affected by the low temperature preservation in biological characteristics.