低温保存对白血病源性树突状细胞的影响.doc

1 低温保存对白血病源性树突状细胞的影响作者: 黄友章, 沈建良, 王立新, 向丹, 郑培浩, 岑坚, 宫立众,刘毅,杨平地【摘要】本研究观察低温保存对白血病源性树突状细胞(L拟 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 拟 DR 、 CD1a 、 CD83 、 CD86 明显上调,而 CD14 表达则明显下调,低温保存前、 2 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 CT L 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拟 DC ;L拟 DC 的诱生在生物学特性方面不受低温保存的影响。【关键词】白血病细胞 Influence of Cryopreservation on Leukemic Dendritic Cells Derived from Leukemia Patients Abstract This study was aimed to investigate the influence of cryopreservation on biological properties and function of leukemic dendritic cells(L 拟 DCs) derived from patients with acute or chronic leukemia. Some fresh leukemic cells were detected immediately; some were cultured immediately; some were cryopreserved in拟 80℃ with 5% DMSO-6% HES as cryopreservor. After bEing thawed , they were cultured. bination of rhGM 拟 CSF, rhIL 拟 4, rhTNF 拟α and other cytokines were added into the culture system. 12 days later, L拟 3 DCs were assayed for morphology, immunophenotype, mixed lymphocytic reaction (MLR) and CTL cytotoxicity on autologous leukemic cells. The results showed that both fresh and cryopreserved leukemic cells obtained from patients with acute or chronic leukemia revealed typical DC morphologically by means of binations of cytokines in culture, but there was no significant difference between pre 拟 or post cryopreservations. L拟 DCs also upregulated the expression of CD80, CD54, HLA 拟 DR , CD1a, CD83 and CD86, and downregulated the expression of CD14, but there was also no difference pared with L拟 DCs befor cryopreservation. L 拟 DCs derived from leukemic cells were also cap