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目的 探讨获得高效肝癌树突状细胞瘤苗(Dendritic Cells,DCs瘤苗)的优化方法。方法 取健康人新鲜血,Ficoll密度梯度离心分离外周血单个核细胞(PBMC),诱导DCs。A组:用重组人粒巨细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)诱导DCs,B组:用GM-CSF和IL-4诱导,肝癌细胞冻融抗原冲击,C组:用GM-CSF、IL-4和肿瘤坏死因子-α(TNF-α)诱导,D组:用GM-CSF、IL-4和TNF-α诱导,肝癌细胞冻融抗原冲击。结果 荧光抗体CD11c和CD54均高表达,各组间无显著性差异(P>0.05),TNF-α可明显上调CD86和CD80表达(P<0.01),肝癌细胞冻融抗原可进一步激活不同条件下诱导的DCs,促进CD86、CD80和人类白细胞抗原-DR(HLA-DR)表达。结论 经GM-CSF、IL-4加TNF-α诱导的DCs可有效的摄取肝癌细胞冻融抗原,促进DCs的成熟,可制备高效的肝癌DCs瘤苗,用于肝癌过继性免疫治疗。
Objective To investigate the optimal method for obtaining dendritic cells (DCs) efficiently. Methods Fresh blood was collected from healthy volunteers. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density gradient centrifugation to induce DCs. Group A: DCs were induced by recombinant human GM-CSF and IL-4; Group B: induced by GM-CSF and IL-4; Group C: induced by GM-CSF, IL-4 and tumor necrosis factor-α (TNF-α); Group D: induced by GM-CSF, IL-4 and TNF-α; Results Fluorescent antibody CD11c and CD54 were all highly expressed (P> 0.05). TNF-α significantly up-regulated the expression of CD86 and CD80 (P <0.01). The freeze-thaw antigen of hepatoma cells could further activate under different conditions Induced DCs, promote the expression of CD86, CD80 and human leukocyte antigen-DR (HLA-DR). Conclusions DCs induced by GM-CSF, IL-4 and TNF-α can efficiently ingest and freeze freeze-thaw antigen of hepatoma cells and promote the maturation of DCs, and can be used to prepare efficient DCs vaccine for hepatocellular carcinoma for adoptive immunotherapy of hepatocellular carcinoma.