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机构地区:[1]第三军医大学基础部全军免疫学研究所,重庆400038
出 处:《现代肿瘤医学》2008年第8期1263-1265,共3页Journal of Modern Oncology
基 金:国家自然科学基金资助项目(编号:30400215)
摘 要:目的:构建基于人乳头状瘤病毒(human papilloma virus,HPV)16型的肿瘤疫苗,并检测其对Hela肿瘤细胞的杀伤活性。方法:利用昆虫杆状病毒表达系统表达病毒蛋白,在体外变性与复性过程中将病毒蛋白包装绿色荧蛋白EGFP和白喉毒素(Diphtheriatoxin,DT)A链(DT-A)的双表达质粒,形成伪病毒疫苗。透射电镜观察病毒样颗粒(virus-like particle,VLP)的结构,并转染Hela肿瘤细胞,荧光显微镜和流式细胞仪检测其转染效率和对Hela肿瘤细胞的杀伤能力。结果:透射电镜观察证实病毒蛋白可自我组装成VLP,在转染Hela肿瘤细胞后,荧光显微镜和流式细胞仪成功检测到荧光的表达和对Hela肿瘤细胞的杀伤作用。结论:基于HPV16的新型伪病毒肿瘤疫苗能成功转染Hela肿瘤细胞并产生杀伤活性,为肿瘤的基因治疗提供了依据。Objective:To construct human papilloma virus type 16 (HPV16) tumor vaccine and to examine its kill activity on Hela cells. Methods: Virus protein was expressed by Bac -to -Bac expression system, EGFP and DT - A double expression plasmid together with virus protein were formed into pseudovirus in the course of degenera- tion and regeneration in vitro. The construction was identified by transmission electron microscope, the transfection and kill activity were detected by fluoroscope and FCM. Results: The pseudovirus could selfassemble into VLP, fluoroscope and FCM could detect the expression of fluorescence and kill activity successfully. Conclusion: Pseud- ovirus tumor vaccine of human papilloma virus type 16 could transfect and kill Hela cells, which might provide a new strategy for tumor gene therapy.
关 键 词:人乳头状瘤病毒16型 肿瘤疫苗 杀伤活性
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