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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 prepare DC vaccine containing hGM-CSF gene and study its antitumor effect. Methods: (1) Umbilical cord blood-derived DCs were isolated and identified by phase-contrast microscopy and flow cytometry; (2) Lenti-hGM-CSF transfected DCs were detected by ELISA, the content of hGM-CSF . (3) DC vaccine stimulated allogeneic T lymphocytes to induce specific CTLs and their proliferation and specific cytotoxicity against target cells were detected by MTT assay. Results: (1) The content of hGM-CSF in DC medium after transfection by Lenti-hGM-CSF was significantly higher than that of untransfected DC cells (P <0.01); (2) Lenti-hGM-CSF transfected cervical cancer DC vaccine stimulated lymphocyte proliferation index (SI) of 2.16, which was significantly better than that of non-transfected cervical cancer DC vaccine. (3) CTL induced by DC vaccine modified by Lenti-hGM-CSF could inhibit cervical cancer HeLa cells The killing activity reached 51.12%, higher than that of unmodified DC induced CTL (P <0.05). CONCLUSION: DC vaccine prepared by Lenti-hGM-CSF modification enhances the ability of stimulating T cell proliferation, and induces CTL to significantly enhance the killing effect on cervical cancer HeLa cells.