目的:构建人乳头状病毒(human papilloma virus,HPV)16型的新型伪病毒,并检测其对DNA特异性B细胞(R4)的靶向性。方法:利用昆虫杆状病毒表达系统表达病毒蚩白,在体外变性与复性过程中将病毒蛋白包装绿色荧蛋白EGFP表达型质粒,形成伪病毒。透射电镜观察病毒样颗粒(virus—like particle,VIJP)的结构,并转染M细胞,荧光显微镜和流式细胞仪检测其转染效率。结果:透射电镜观察证实病毒蛋白可自我组装成VLP,在转染R4A细胞后,荧光显微镜和流式细胞仪成功检测到荧光的表达。结论:HPV16的新型伪病毒能成功转染M细胞,为系统性红斑狼疮(SLE)的靶向治疗提供了理想的策略。
Objective:To construct pseudovirus of human papilloma virus type 16 (HPV16),and to examine its target activity on DNA specific B cells (R4A). Methods:Virus protein was expressed by Bac-to-Bac expression system, EGFP expression plasmid together with virus protein were formed into pseudovirus in the course of degeneration and regeneration in vitro.The construction was identified by transmission electron microscope,the transfection and target activity were detected by fluoroscope and FCM.Results:The pseudovirus could selfassemble into VLP,fluoroscope and FCM could detect the expression of fluorescence successfully. Conclusion:Pseudovirus of human papilloma virus type 16 could transfect R4A cells, which might provide a new strategy for SLE theray.