人乳头瘤病毒(HPV)L2肽免疫小鼠诱导的抗体水平与保护作用的关系研究

摘要

Objective To detect the protection induced by HPV-58 L2 11-200 AA in animal, and analyze the relationship between antibody or neutralizing antibody titers and the protection generated by the immunizmg agent. Methods The peptide of HPV-58 L2 11-200 AA was expressed in E. coli and the mice were immunized with the peptide after purification and adsorption with aluminum adjuvant. The protection provided by different immunizing doses was detected in the mouse model against the challenge of the pseud-ovirions of human papiilomavirus types 58. The total antibodies and neutralizing antibody titers of serum were tested with ELISA and neutralization assay against HPV-58 pseudovirus, respectively. The total antibodies or neutralizing antibody titers that can protect the mouse from infection were analyzed. Results The mice can be protected from the challenge with HPV pseudovirus when the immunizing dose was 8 μg. The neutralizing antibody can not be detected in the immune serum by neutralization assay against pseudovirus. The total anti-body level has a corresponding relationship with the protection showed in mouse model. The results of total antibodies detected by ELISA showed that when the titer of total antibodies was ≥25 000, luminescent signal can not be detected and the mice can be protected from pseudovirus infection. Conclusion HPV-58 L2 11-200 AA peptide can protect mice from pseudovirus infection. L2 peptide has a promising perspective to be a candidate vaccine and the level of total antibodies in the immune serum can be used as a surrogate for the evaluation of protection against HPV infection.%目的 检测HPV-58 L2 11～200 AA肽在动物体内对HPV的保护效果,并分析疫苗诱导的抗体滴度或中和抗体滴度与疫苗保护作用之间的对应关系.方法 利用大肠杆菌表达HPV-58L2 11～200 AA肽,纯化目的 蛋白并与铝佐剂吸附后免疫小鼠,利用小鼠HPV-58假病毒感染模型检测不同剂量的免疫原对小鼠的保护作用.通过ELISA和假病毒中和抗体检测方法 检测免疫血清中的总抗体水平和中和抗体水平,分析具有保护作用的抗体或中和抗体滴度.结果 当蛋白免疫剂量为8μg时,能够完全保护小鼠不受HPV假病毒的感染.小鼠免疫血清中的中和抗体水平较低,利用已建立的假病毒中和试验方法 检测不到血清中的中和抗体.而利用ELISA检测血清中的总抗体水平结果 显示,免疫血清中的总抗体水平与疫苗的保护效果之间存在对应关系,当抗体滴度大于等于1:25 000时,小鼠体内检测不到荧光信号,能够保护机体不受假病毒的感染.结论 HPV-58 L2 11～200 AA肽能够保护小鼠不受假病毒的感染,L2 11～200 AA肽具有较好的疫苗发展前景,其引发的总抗体水平可以作为评价保护效果的间接指标.