肺癌细胞总RNA转染的DC疫苗体外诱导肺癌患者特异性抗肿瘤免疫的实验研究

Experimental study of specific antitumor immunity induced by dendritic cell vaccine transfected with tumor cell total RNA in lung cancer patients

目的 :为了探讨肺癌细胞总RNA转染的DC疫苗体外诱导特异性抗肿瘤免疫的能力。方法 :采用分离肺癌患者外周血单核细胞体外诱导DC细胞 ,Trizol法提取肺癌细胞系Calu 6总RNA ,用脂质体包裹总RNA转染DC并诱导特异性CTL的扩增 ,LDH法和ELISA法检测CTL的杀伤活性和IFN γ分泌。结果 :经肺癌细胞总RNA转染的DC特异性表面标志及功能相关分子表达均上调 ,转染后的DC可显著刺激异体 自体T淋巴细胞增殖 ,诱导的特异性CTL对携带Calu 6抗原的靶细胞的杀伤率显著高于LAK细胞 ,再次接触相同抗原时其IFN γ分泌量显著增高。结论 :肺癌细胞总RNA转染的DC疫苗可在体外诱导出特异性抗肿瘤免疫。

Objective:To observe ability of DC vaccine transfected with tumor cell total RNA to induce specific antitumor immunity in lung cancer patients in vitro.Methods:DCs were generated from lung cancer patients' peripheral blood mononuclear cells(PBMC).Total RNA was isolated from lung cancer tumor cell line Calu-6 by Trizol.Autologous DCs transfected with Calu-6 total RNA by liposome were used to induce specific CTL proliferation.Specific cytotoxicity and IFN-γ secretion were measured by LDH assay and ELISA method.Results:Transfected DCs exhibited dramatically increased expression of specific membrane markers and function-associated molecules,and were more potent in stimulating allogenous or autologous T cell proliferation than that of control DCs.Specific CTLs induced by transfected DCs showed higher cytotoxicity than LAK against Calu-6 antigens positive target cells.When sensitized lymphocytes were restimulated by transfected DCs again,IFN-γ secretion enhanced significantly.Conclusion:Lung cancer patient's autologous DCs transfected with tumor total RNA are effective vaccines in stimulating specific antitumor T-cell immunity in vitro.