基于HPV16的新型伪病毒对DNA特异性B细胞的杀伤研究

目的构建基于人乳头状病毒（humanpa pilloma virus,HPV）16型的新型伪病毒,并检测其对DNA特异性B细胞（R4A）的杀伤效应。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B细胞特异性启动子Igκ控制下的白喉毒素（Diphtheriatoxin,DT）A链（DT-A）基因表达型质粒DNA,形成伪病毒。转染R4A细胞,荧光显微镜和流式细胞仪检测其杀伤效应,酶联免疫法检测R4A细胞分泌抗ds-DNA抗体的滴度。结果转染R4A细胞48h后,荧光显微镜和流式细胞仪成功检测到新型伪病毒对R4A细胞的杀伤效应,R4A细胞分泌抗ds-DNA抗体的滴度明显降低。结论基于HPV16的新型伪病毒能有效特异杀伤R4A细胞,并降低抗ds-DNA抗体的滴度,为系统性红斑狼疮的治疗提供了理想的策略。

Study on kill effect of pseudovirus of human papilloma virus type 16 on DNA specific B cells

Objective To construct pseudovirus of human papilloma virus type 16 （HPV16） and to examine its kill activity on DNA specific B cells（ R4A）. Methods Virus protein was expressed by Bac-to-Bac expression system,DT-Aexpression plasmid together with virus protein were formed into pseudovirus in the course of degeneration and regeneration in vitro. R4A cells were transfected with pseudovirus,and kill activity was detected by fluoroscope and FCM. The titer of ds-DNA antibody was also detected by ELISA assay. Results After 48h of transfection,fluoroscope and FCM could detect the kill activity successfully,and the titer of ds-DNA antibody decreased obviously. Conclusion Pseudovirus of human papilloma virus type 16 could specifically kill R4A cells and control the lever of ds-DNA antibody,which might provide a new strategy for SLE theray.