白细胞介素-12基因修饰对树突状细胞表面分子表达及细胞因子分泌的影响

目的 观察白细胞介素(IL)-12基因修饰对树突状细胞(DC)表面分子及细胞因子分泌的影响.方法 采用重组逆转录病毒介导IL-12基因修饰人外周血单个核细胞(PBMC)来源的Dc;ELISA法检测各组DCs和各组T细胞上清中IL-12、IL-10、IFN-γ因子的分泌水平;流式细胞仪(FACS)分析各组DC表面CD83、CD86的表达;MTT法检测DC刺激同源T淋巴细胞增殖的能力;统计学分析比较各组间的差异.结果 IL-12基因修饰使得DC高表达CD83和CD86分子,分泌高水平IL-12及IFN-γ,但对IL-10因子的分泌无明显影响,刺激同源T淋巴细胞增殖明显,诱导激活的T细胞上清中IFN-γ水平显著增高、IL-10分泌水平显著降低.结论 经IL-12基因修饰后的DC表型成熟,分泌IL-12及IFN-γ的能力增强,对IL-10因子的分泌无影响,能抑制T细胞分泌IL-10因子,优化抗原提呈的微环境.

Effect of IL-12 gene modification on the phenotype and cytokine secretion of dendritic cells

Objective To investigate the effects of interleukin-12 gene modification on the phenotype and cytokine secretion of dendritic cens(DCs)in vitro.Methotis DCs were derived from human peripheral blood mononudear cells(hPBMCs).modified bv recombinant IL-12 retrovirus.The IL-12,IL10 and IFN-γ levels in culture supematant of DCs and T lymphocytes were examined by ELISA.The expression of CD83 and CD86 molecules on DCs surface was detected bv FACS.Proliferation capacity of activating autologous T lymphocytes was evaluated by MTT.Results After being modified by IL-12 gene.DCs produced high level of IL-12 and IFN-γ but had on effect on the production of IL-10,and increased the expression of CD83 and CD86 molecules.Proliferation of autologous T lymphocytes was also enhanced after being stimulated by DC vaccine.DC vaccine could induce and activate T lymphoeytes to produce high level of IFN-γ,but suppress the production of IL-10.Conclusion After being modified by IL-12 gene,DCs showed more mature.