摘要
目的建立人乳头状瘤病毒（HPV）假病毒小鼠感染模型。方法将密码子优化的HPV衣壳蛋白基因表达质粒和荧光素酶报告基因表达质粒共转染293FT细胞，48h后收集裂解细胞，收获假病毒，并对假病毒滴度进行测定。试验小鼠皮下注射甲羟孕酮醋酸酯，4d后阴道灌注壬苯醇醚-9，并在6h后阴道灌注假病毒，7d后阴道灌注荧光素酶底物，并检测荧光素酶报告基因的表达情况。结果成功制备了3种型别（HPV16、HPV45和HPV58）的假病毒，其滴度分别为3．7×10^8TU／ml、1．5×10^8TU／ml和1．2×10^8TU／ml。在3种假病毒感染的小鼠体内可见发光区域，其荧光信号强度分别为1．779×10^6p／s、5．738×10^5p／s和1．829×10^6p／s。结论成功建立了HPV16、45和58型的小鼠感染模型，为HPV感染干预研究、疫苗评价及预防药物的筛选奠定基础。
Objective To establish the mouse model for human papillomavirus types 16, 45 and 58 by the corresponding pseudovirions. Methods The 293VF cells were co-transfected with codon-modified HPV capsids genes together with a reporter plasmid containing the luciferase gene. The cells were collected and lysed, then the pseudovirus was collected and the titration was performed. The mouse was subcutaneously injected with Depo-Provera. After 4 d and intravaginally injected with nonoxynol-9, and 6 h later pseudoviruses were inoculated in intravaginal. After 7 d, the mouse was instilled luciferin substrate intravaginally, and the expression level of the luciferase gene was detected by the in vivo Imaging System （IVIS）. Results Three types （ HPV16, HPV45 and HPV58 ） of pseudnviruses had been produced and the titer was 3.7 ×10^8 TU/ml, 1.5 ×10^8 TU/ml and 1.2 ×10^8 TU/ml, respectively. The luminescent regions could be detected in the mice which were infected with the pseudovirions, the luminescent signal intensity for types 16, 45 and 58 was 1. 779 ×10^6 p/s, 5. 738 ×10^5 p/s and 1. 829 ×10^6 p/s, respectively. Conclusion The mouse models for HPV16, 45 and 58 have been successfully established based on pseudovirions, which will be very useful for the research of HPV infection intervention, the evaluation of HPV vaccines and the screening of the prophylactic agents.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2008年第9期791-794,共4页
Chinese Journal of Microbiology and Immunology
