SOCS1siRNA和IL-12基因共同修饰树突状细胞体外抗喉癌的免疫反应

目的:探讨腺病毒负载SOCS1siRNA和IL-12基因共同修饰人喉癌细胞(Hep-2)抗原致敏的树突状细胞(DC)在体外诱导﹑激活细胞毒性T淋巴细胞(CTL)免疫杀伤Hep-2的效能及相关免疫机制。方法:采用重组腺病毒介导SOCS1siRNA基因和重组腺病毒介导IL-12基因共同修饰人外周血单个核细胞(PBMC)来源的DC;采用反复冻融法提取Hep-2粗体抗原(Ag)致敏的基因修饰的DC;用ELISA法检测各组DC及CTL分泌IL-12和IFN-γ的水平;MTT法检测DC刺激同源T淋巴细胞增殖能力及CTL免疫杀伤Hep-2的效能;统计学分析各组间的差异。结果:DC体外诱导培养成功,Ad-GFP转染可见荧光表达率在90%以上。经SOCS1siRNA和IL-12基因共同修饰能有效下调DC中SOCS1蛋白的表达和上调IL-12蛋白的表达。IL-12因子的分泌率也明显高于SOCS1siRNA或IL-12的单基因转染,并能促使T细胞显著增殖和活化。DC和CTL持续高分泌IFN-γ,因此产生对Hep-2的特异性细胞免疫。结论:SOCS1siRNA和IL-12基因共同修饰喉癌抗原致敏的DC分泌的IL-12和IFN-γ的能力增强。双基因共同修饰喉癌抗原致敏的DC与T细胞混合反应能有效的促进T细胞增殖,促进其分泌IFN-γ和IL-12,从而诱导CTL更强的特异性杀伤喉癌细胞的能力。

Silencing of SOCS1and IL-12 gene cotransfered by adenoviral enhances DC-mediated anti-laryngocarcinoma immunity in vitro

Objective:To investigate the effects and the related immunological mechanisms of dendritic cells(DCs) modified by SOCS1siRNA gene and IL-12gene on activating and inducing cytotoxic T lymphcyte(CTL) as well as specific immunitically killing tlaryngocarcinoma in vitro.Method:DCs were derived from human peripheral blood mononuclear cells(hPBMC),modified by recombinant SOCS1siRNA adenoviral and IL-12adenoviral and then pulsed with tumor antigen of repeated freeze-thaw method.The IL-12and IFN-γlevels in culture supernatant of DCs and CTLs were examined by ELISA.Result:DC were cultivated successfully and had special morphologic haracteristicistics.The rate of Ad-GFP carrying fluorescent expression was over 90%.The expression of SOCS1 protein in DCs were effectively decreased by being modified SOCS1siRNA and IL-12genetic while the expression of IL-12protein were increased.The secretion rate of IL-12factor was higher than that of SOCS1siRNA and IL-12 transfection of single gene respectively in modified DCs which could prompt T cell proliferation activation significantly as well.IFN-γwas secreted constantly in DC and CTL,resulting in Hep-2.Conclusion:DC modified by SOCS1siRNA and IL-12gene which plused with larygneal carcinomal antigene could increased the production of IL-12and IFN-γ;DC modified by SOCS1siRNA and IL-12gene which plused with larygneal carcinomal antigene could enhance the ability to stimulate proliferation of T cell,increase production of IFN-γ,IL-12by T cells and induce the stronger killing rate of CTL.