中国免疫学杂志2011，Vol.27Issue(2)：115-119,5.DOI:10.3969/j.issn.1000-484X.2011.02.005
HPV 16 E6E7基因修饰树突状细胞疫苗诱导CTL致CaSki细胞凋亡体外实验研究
Construction of genetically modified dendritic cell vaccine expressing HPV-16 E6E7 fusion gene and activating specitific cytotoxic T lymphocytes to induce apoptosis CaSki cells in vitro
摘要
Abstract
Objective:To develop the recombinant adenovirus carrying HPV-16 E6E7 (pAd-E6E7) to infect mouse immature dendritic cells, and to observe the apoptosis of CaSki cells induced by specific cytotoxic T lymphocytes (CTL) triggered by genetically modified DC vaccine expressing HPV-16 E6E7 fusion gene in vitro.Methods:The pAd-E6E7 was used to infect mouse immature dendritic cells to prepare DC vaccine, the confocal microscopy was used to observe the expression of green fluorescence protein in immature dendritic cells, and flow cytometry (FCM) was used to examine the surface expression of CD40, CD86, MHC Ⅱ and CDUC in both uninfected and infected dendritic cells.DC vaccine was used to induce specific CTL,and after co-culturing the CTL with CaSki cells, DAPI,TUNEL and flow cytometry (FCM) were used to examine the apoptosis of CaSki cells.Results: Immature mouse dendritic cells were successfully transfected by pAd-E6E7 in vitro, and the transfecting efficiency was 40％-50％.DC vaccine was successfully prepared and used to induce specific CTL.CaSki cells underwent apoptosis via coculturing with the specific CTL detected by DAPI,TUNEL and flow cytometry (FCM).Conclusion:Genetically modified DC vaccine expressing HPV-16 E6E7 fusion gene could induce CTL, and the CTL could induce apoptosis of CaSki cells in vitro.关键词
人乳头状瘤病毒/细胞毒性T淋巴细胞/CaSki细胞/树突状细胞分类
医药卫生引用本文复制引用
焦庆昉,易发平,汪长东,袁成福,刘勒,兰欢,符少月,艾青,宋方洲..HPV 16 E6E7基因修饰树突状细胞疫苗诱导CTL致CaSki细胞凋亡体外实验研究[J].中国免疫学杂志,2011,27(2):115-119,5.基金项目
本文受国家自然科学基金项目(30800945)和重庆市科委自然科学基金项目(CSTC,2008BB5225)资助 （30800945）
