摘要
目的探讨转染IL-27基因体外对人外周血单个核细胞来源树突状细胞表型和功能的影响。方法采集健康成人外周血,密度梯度离心法获得外周血单个核细胞,用GM-CSF、IL-4诱导树突状细胞(Dendritic cell,DC),第5天后将DC分为3组:IL-27基因转染组、TNF-α组和阴性对照组。倒置显微镜观察DC形态;流式细胞术检测DC表面分子CD1a、CD80、CD83和CD86的表达情况;MTT法检测DC刺激T细胞增殖的能力;ELISA法检测细胞培养上清中IL-12和IFN-γ的含量。结果转染IL-27基因后外周血单个核细胞来源DC呈现典型的成熟DC形态学特征。IL-27基因转染组DC表面CD1a、CD80、CD83和CD86表达水平较阴性对照组均明显上调(P<0.05)。IL-27基因转染组DC诱导T细胞增殖的能力较对照组DC明显增高(P<0.05)。IL-27基因转染组DC培养上清中IL-12和IFN-γ的含量均明显高于对照组DC(P<0.01)。结论转染IL-27基因可以上调成熟DC的细胞表型,增强DC的免疫学活性。
Objective To investigate the effects of IL-27 gene on phenotype and function of DCs（Dendritic cell） derived from PBMCs（peripheral blood mononuclear cells）.Methods PBMCs isolated from healthy human peripheral blood were cultured with GM-CSF（granulocyte-macrophage clone stimulating factor）plus IL（Interleukin）-4 for 5 days,and then were divided into three groups：IL-27 gene transfection,TNF（tumor necrosis factor）-α and control group.The morphological characteristics of DCs were observed by inverted microscope.Expressions of CD1a,CD83 and CD80,and CD86 on DCs were analyzed with flow cytometry.MTT method was used to determine the proliferation of T cells stimulated by DCs.ELISA was used to detect the production of IL-12 and IFN-γ in the culture supernatant of DCs.Results PBMCs were developed into mature DCs with typical morphological characteristics after IL-27 gene transfection.Expressions of CD1a,CD83 and CD80,and CD86 on DCs in IL-27 gene transfection and TNF-α groups were up-regulated compared to the control group（P0.05）.IL-27 gene transfected DCs markedly stimulated the proliferation of T cells and enhanced the production of IL-12 and IFN-γ（P0.05）.Conclusion IL-27 gene transfection could up-regulate the phenotype of mature DCs and enhance the immune activity of DCs.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2011年第1期31-34,共4页
Cancer Research on Prevention and Treatment
基金
河北省科技厅基金资助项目(10276105D-98)
河北省卫生厅基金资助项目(20100413)
