重组质粒pchIL-18-MAGE转染树突状细胞疫苗体外杀伤肝癌细胞的研究

Study of killing effect on liver cancer cells by transfected dendritic cell vaccine of recombinant plasmid pch IL-18-MAGE in vitro

目的:探讨重组质粒pch IL-18-MAGE转染DC细胞在体外对肝癌细胞的杀伤作用。方法:构建共表达质粒pchIL-18-MAGE,体外培养树突状细胞,将以上重组质粒转染树突状细胞。RT-PCR和Western blot方法验证IL-18和MAGE-1基因在转染DC中的表达。应用流式细胞术检测转染后DC细胞的表型变化。以转染重组质粒DC刺激的淋巴细胞作为效应细胞,单纯淋巴细胞和未经转染DC刺激的淋巴细胞为对照,检测其对肝癌靶细胞的体外杀伤作用。ELISA法检测INF-γ的分泌。结果:pchIL-18-MAGE转染后DC细胞高表达CD83、CD1a、CD86、CD80、HLA-DR等抗原,表现为成熟DC表型特征。共表达质粒转染的DC细胞诱导的CTL对肝癌细胞杀伤作用最强(P<0.05)。结论:pchIL-18-MAGE转染DC细胞对MAGE+肝癌细胞杀伤作用明显。

Objective：To observe lethal effect on liver cancer cells by the DCs transfected with recombinant plasmid pchIL -18-MAGE.Methods：Cultivate dendritic cells in vitro ,and recombinant plasmid pchIL-18-MAGE was constructed which was transfected into DCs.Expression of IL-18 and MAGE-1 on the transfected DCs were detected by RT-PCR and Western blot.The diversity of DCs phenotypes after transfect were analyzed by flow cytometry.Different killing effects to the target cells by lymphocytes which were stimulated by DC and those by non-transfected DC were detected.IFN-γin the cellular supernatant of the transfected DCs was inspected by ELISA.Results：pchIL-18-MAGE transfected DCs expressed higher level of CD 83,CD1a,CD86,CD80,HLA-DR,and expressed as a mature DC phenotype.pchIL-18-MAGE group′s killing activity was the highest （ P〈0.05 ） .Conclusion： The DC vaccine pchlL-18-MAGE can induce better killing effect compared with other experimental groups.