目的: 研究流感疫苗对髓系白血病骨髓源性树突状细胞（dendritic cells，DCs）功能的影响及其机制。方法：分离髓系白血病患者\[急性髓细胞白血病(acute myeloid leukemia, AML)19例， 慢性髓细胞白血病（chronic myeloid leukemia，CML) 8例\]骨髓单个核细胞（mononuclear cell，MNC），用GMCSF和IL4诱导7 d，获得未成熟白血病DCs，然后加入全病毒灭活流感疫苗（whole inactivated influenza vaccine, WIV）、裂解病毒流感疫苗（split influenza vaccine，SIV）或TNFα继续培养24 h。R显带法分析DCs染色体核型，流式细胞仪检测DCs表型，ELISA法测定DCs培养上清IL12的水平，CCK8法检测DCs诱导的CTL对自体白血病细胞的细胞毒作用。结果：19例AML患者中的15例及8例CML患者的MNC全部成功诱导出DCs。与TNFα刺激的白血病DCs相比，流感疫苗刺激的白血病DCs表面分子（CD80、CD83、CD86、HLADR）表达明显上调（P<005），培养上清中IL12的分泌水平明显增加（P<0.05），其诱导的CTL可显著杀伤自体白血病细胞（P<0.05）；WIV刺激的DCs在表型、IL12分泌水平及细胞毒作用方面均较SIV刺激的DCs显著增高（P<0.05）。结论： 流感疫苗促进髓系白血病源DCs表型成熟及IL12的分泌，增强其诱导的CTL对自体白血病细胞的杀伤作用。

Objective:To determine whether the influenza vaccine can affect the function of dendritic cells（DCs）derived from the bone marrow of patients with myeloid leukemia and the possible mechanism. Methods: The bone marrow (BM) mononuclear cells were obtained from 19 patients with acute myelocytic leukemia (AML) and 8 patients with chronic myeloid leukemia (CML), and were cultured with GMCSF and IL4 for 7 days to obtain immature DCs. Then DCs were stimulated by whole inactivated influenza vaccine (WIV), split influenza vaccine (SIV), or TNFα. After 24 h, phenotypes and karyotypes of these DCs were assessed by FACS and R band karyotype analysis, respectively. The supernatant IL12 levels were measured by ELISA in each group. Cytotoxic activity of CTL induced by differently treated DCs was measured by CCK8 assay.Results: DCs were successfully induced in 15 of the 19 AML patients and all the 8 CML patients. After stimulated with WIV or SIV for 24 h, DCs exhibited enhanced expression of CD83, CD86 and HLADR, and increased secretion of IL12 (all P<0.05). CTL induced by WIV or SIVstimulated DCs specifically killed autologous leukemia cells in vitro (P<0.05). Furthermore, WIVstimulated DCs were more powerful than SIVstimulated DCs in killing target cells (P<0.05). Conclusion: Influenza vaccine can promote the maturation and IL12 secretion of DCs derived from myeloid leukemia patients, and CTL induced by influenza vaccinestimulated DCs has a stronger ability to kill autologous leukemia cells.

甘肃省科学基金资助项目（No.0709TCYA063）