负载hGM-CSF基因的慢病毒载体介导的宫颈癌DC疫苗抗肿瘤活性的初步研究

Anti-tumor effects of lentivirus-hGM-CSF mediated cervical cancer DC vaccine

目的:制备负载hGM-CSF基因的宫颈癌DC疫苗并研究其抗肿瘤作用。方法:(1)分离制备脐血来源的DC,通过相差显微镜和流式细胞仪鉴定;(2)Lenti-hGM-CSF转导DC细胞,ELISA方法检测DC细胞上清液中hGM-CSF的含量。宫颈癌HeLa细胞冻融抗原刺激DC细胞制备DC疫苗;(3)DC疫苗刺激同种T淋巴细胞诱导特异性CTL并用MTT法检测其增殖能力和对靶细胞的特异性杀伤能力。结果:(1)经Lenti-hGM-CSF转导后DC培养液中hGM-CSF含量明显高于未转染的DC细胞(P<0.01);(2)Lenti-hGM-CSF转染的宫颈癌DC疫苗刺激淋巴细胞的增殖指数(SI)达2.16,明显优于未转染的宫颈癌DC疫苗;(3)经Lenti-hGM-CSF修饰的宫颈癌DC疫苗诱导的CTL对宫颈癌HeLa细胞的杀伤活性达51.12%,高于未经修饰的DC诱导的CTL(P<0.05)。结论:用Lenti-hGM-CSF修饰制备的宫颈癌DC疫苗,其刺激T细胞增殖的能力增强,诱导的CTL对宫颈癌HeLa细胞具有显著增强的杀伤效应。

Objective：To produce gene modified cervical cancer DC vaccine transduced with lentiviral vector encoding hGM-CSF and to investigate its anti-tumor effect. Methods：（ 1 ）Dendritic cells from cord blood were isolated and induced, their morphology and phe- notype were analyzed by flow cytometer. （2）Dendritic cells were transduced with Lenti-hGMCSF and were charged with HeLa cell lysate to get gene modified DC vaccine. The concentration of hGM-CSF in DC supernatant was determined by ELISA. （3）The proliferation of allogeneic T lymphocytes stimulated by and the cytotoxicity of CTL activated by Lenti-hGM-CSF DC were detected with MTT assay. Results： （ 1 ） Lenti-hGM-CSF DC secreted more hGM-CSF than did DC control groups（P〈0.01 ）. （2）The stimulatory capacity of the Lenti-hGM-CSF DC in the allogeneic T lymphocytes reaction was markedly enhanced as compared with DC control groups（P 〈0.05）. The stimulator index （SI） were all above 1.5 in Lenti-hGM-CSF DC group. （3） The killing activity of CTL stimulated with Lenti-hGM-CSF-DC to HeLa was significantly higher than that of CTL stimulated with DC control groups （P〈0.05）, and the killing rate was 51.2%. Conclusion：The gene modified DC vaccine transduced with lenti-hGM-CSF can stimulate the proliferation of allogeneic T lymphocytes more effectivly, and induce CTL to kill HeLa more efficiently.