肝癌细胞冻融抗原制备树突状细胞瘤苗的研究

The using hepatocellular carcinoma cell lysate antigen to produce DCs Vaccination

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 obtain the method of highly generative Dendritic cells vaccination of hapa-tocellular carcinoma from peripheral blood. Methods The PBMC were islated from the healthy donors by Fi-coll. The adherent PBMC were harvested and induced to be DCs. The cells were divided into different groups. Group A: DCs were stimulated by rhGM-CSF and IL-4; Group B: DCs were stimulated by rhGM-CSF, IL-4 and lysate hepatocellular carcinoma cell; Group C: DCs were stimulated by rhGM-CSF, IL-4 and TNFα; Group D: DCs were stimulated by rhGM-CSF, IL-4, TNF-α and lysate hepatocellular carcinoma cells. Results DCs highly expressed CD11 and CD54( P > 0. 05) in all groups. TNFa could significantly stimulate the DCs to express CD80 and CD86 (P < 0. 01), Lysate hepatocellular carcinoma cells soulds induce DCS to express CD86 ( P < 0. 01). Lysate hepatocellular carcinoma cells could induce DCs to express CD86, CD80 and HLA-DR in different conditions. Conclution DCs stimulated by GM-CSM, IL-4 and TNF-α can accept the antigen from hepatocellular carcinoma cells, it can be used as vaccination for HCC patients.