同济大学学报（医学版）2012，Vol.33Issue(1)：13-18,23,7.DOI:10.3969/j.issn1008-0392.2012.01.003
靶向树突状细胞的肿瘤疫苗OVA慢病毒载体构建及其体外抗肿瘤效应
Construction of ientiviral vector targeting dendritic cell vaccine and its antitumor effects
摘要
Abstract
Objective To construct lentiviral vector containing simulated tumor antigen ovalbumin (OVA) targeting dendritic cell(DC) vaccine and to test its antitumor effects in vitro. Methods The lentiviral expression vector pHR-CMV-EGFP-OVA was constructed by introducing DNA fragment of OVA into the Xho I and Xba I sites of pHR-CMV-EGFP vector. DNA sequencing and restriction endonucleases methods were used to test the recombinant vector. DCs were collected and isolated from the mouse bone marrow and purified by FACS. 293 T cells were transfected with the expression vector pHR-CMV-EGFP-OVA by liposome-medium transfection, and then Lewis lung carcinoma (LLC) cells were transfected with pHR-CMV-EGFP-OVA lentivirals. The positive clones were expanded and screened for OVA production by RT-PCR and Western blot. DCs also were transfected with pHR-CMV-EGFP-OVA. T cells were collected from spleen and isolated by using Nylon wool column. Results The lentiviral vector pHR-CMV- EGFP-OVA was constructed successfully. LLC cells and DCs carrying ovalbumin were successfully obtained. Tumor-specific T cells stimulated by OVA-loaded DCs exhibited effectively cytolytic activity against LLC/OVA cells in vitro. Conclusion The lentiviral expression vector pHR-CMV-EGFP-OVA has been successfully constructed and the transfected DCs vaccine can elicit the anti-tumor immunity to OVA-loaded Lewis lung cancer cells in vitro.关键词
慢病毒载体/肿瘤抗原/树突状细胞/肺肿瘤细胞/肿瘤免疫治疗/基因治疗Key words
lentiviral vector/ tumor antigen/ dendritic cell/ lung cancer cell/ tumor immunother-apy/ gene therapy分类
医药卫生引用本文复制引用
陈帅,李浩博,李牧,杨耀琴,陶惠红,赵培林..靶向树突状细胞的肿瘤疫苗OVA慢病毒载体构建及其体外抗肿瘤效应[J].同济大学学报（医学版）,2012,33(1):13-18,23,7.基金项目
上海市教委科研创新项目重点项目(08ZZ19) （08ZZ19）
