K562细胞和树突状细胞在细胞因子表达方面的相互作用

The interaction between K562 cells and dendritic cells on the expression of cytokines

目的:探讨共培养条件下血液肿瘤K562细胞和树突状细胞(dendritic cells,DCs)在表达细胞因子方面的相互作用情况,探索血液肿瘤发生发展过程中的免疫逃逸机制。方法:利用免疫磁珠试剂盒从人健康外周血中分离纯化出高纯度的CD14+单核细胞,一定浓度的巨噬细胞集落刺激因子(granlocyte-macrophage colomy stimulating factor,GM-CSF)和白介素4(interlenkins-4,IL-4)将单核细胞诱导分化为未成熟DCs(immature DCs,im DCs),再用一定浓度的肿瘤坏死因子(tumor necrosis factor-α,TNF-α)将im DCs诱导为成熟DCs(mature DCs,m DCs),分别将im DCs和m DCs与K562细胞在Transwell系统中共培养48h,对照组为正常培养48h的DCs和撤细胞因子培养48h的DCs,ELISA方法分别检测各组细胞上清液中IL-10、IL-12、转化生长因子β1(transformed growth factor-β1,TGFβ1)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达情况。结果:共培养后,DCs表达细胞因子IL-12减少,DCs和K562细胞表达TGFβ1增加,K562细胞表达VEGF增加。结论:共培养后,两种细胞在某些机制的相互作用下,DCs表达IL-12能力降低,DCs和K562细胞表达TGFβ1的能力增高,K562细胞表达VEGF的能力提高,这可能是共培养后DCs免疫功能下降和血液肿瘤细胞逃脱机体免疫监视的细胞因子方面的原因。

Objective：To study the interactions of dendritic cells（DCs）and K562 when co -culture on cytokines expression,to investigate the mechanisms of leukemia immune escape.Methods：The CD14 ＋ monocytes were purified from fresh peripheral blood of healthy human with immune magnetic beads,and then were cultured in the media con-taining certain concentration rhGM-CSF and IL -4 main to induce into immature DCs（imDCs）,and mature DCs（mDCs）were developed by add certain concentration of rhTNF -αinto imDCs culture.The imDCs and mDCs were co -cultured with K562 cells in transwell chamber for 48h,those untreated cells including those cultured in normal medium for 48h and those cultured in medium without cytokines for 48h served and K562 cells cultured in 1640 for 48h as controls.The expression of IL -10,IL -12,transformed growth factor -β1 （TGFβ1 ）and vascular endothelial growth factor（VEGF）were measured by ELISA.Results：After co -cultured,the expression of IL -12 of DCs were decreased（P 〈0.01）,the expression of TGFβ1 of DCs and K562 cells were increased deeply （P 〈0.01）and the ex-pression of VEGF of K562 cells were increased obviously.Conclusion：After co -cultured,under the interaction of DCs and K562 cells in some mechanism,DCs ability of expressing IL -12 is reduced,DCs and K562 cells ability of expressing TGFβ1 were higher,and K562 cells ability of expressing VEGF were higher too,this could be the reason for DCs immune function damaged and K562 cells to escape immune surveillance on cytokine secretion.