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目的 研究重组人Flt3配体 (rhFL)及其他细胞因子在体外对免疫细胞的抗辐射作用及其可能机理。方法 用流式细胞仪分析照射前后外周血单个核细胞 (PBMC)表型及rhFL对受照射PBMC表型的影响。 [3H] TdR测定细胞的增殖能力。PBMC来源贴壁细胞经细胞因子诱导树突状细胞 (DC) ,并观察辐射对诱导DC的影响及rhFL对DC的抗辐射效应 ;用PI AnnexinV、DNALadder分析细胞凋亡。结果 PBMC在体外经 15Gy照射后 ,CD8+ 、CD5 6 + 细胞明显减少 ,照射前细胞培养液中加入 10 0ng mlrhFL ,可促进CD5 6 + 细胞的恢复 ;照射前应用rhFL及照射后联合应用IL 2等细胞因子 ,可促进受照射CD8+ 、CD5 6 + 细胞的恢复 ;rhFL可协同GM CSF、IL 4、TNFα产生DC ,并使DC细胞的数量较常规方法诱导产生的数量增加 6倍 ,而且rhFL对DC分化诱导的辐射抑制作用具有明显的抗性 ;rhFL可抑制照射所致的细胞凋亡。结论 rhFL单独或与其他细胞因子联合应用 ,对人外周血免疫细胞具有明显的抗辐射效应 ,这对辐射损伤机体免疫功能的调节和恢复具有重要的使用价值
Objective To study the anti-radiation effect of recombinant human Flt3 ligand (rhFL) and other cytokines on immune cells in vitro and its possible mechanism. Methods Flow cytometry was used to analyze the phenotype of peripheral blood mononuclear cells (PBMC) and the effect of rhFL on the phenotype of irradiated PBMC. [3H] TdR was used to measure the proliferation of cells. Dendritic cells (DCs) were induced by cytokines in adherent cells of PBMCs. The effects of radiation on induced DCs and the anti-radiation effects of rhFL on DCs were also observed. Apoptosis was analyzed by PI Annexin V and DNALadder. Results After PBMC was irradiated with 15Gy in vitro, the number of CD8 + and CD5 + cells decreased obviously. The addition of 10ng mlrhFL to the cell culture medium before irradiation could promote the recovery of CD56 + cells. The application of rhFL before irradiation and the combination of IL2 Cytokines can promote the recovery of irradiated CD8 + and CD5 6 + cells; rhFL can produce DC in combination with GM CSF, IL 4 and TNFα, and increase the number of DC cells by 6 times than the conventional method, Differentiation-induced radiation inhibition has obvious resistance; rhFL can inhibit irradiation-induced apoptosis. Conclusion rhFL, alone or in combination with other cytokines, has obvious anti-radiation effects on immune cells of human peripheral blood, which has important value in the regulation and recovery of immune function of radiation-injured organism