一种可诱发高滴度的针对人乳头瘤病毒HPV16,HPV52,HPV58中和抗体反应的HPV16嵌合病毒样颗粒疫苗的研究

A Human Papillomavirus Type 16 Chimeric Virus-like Particle Which Displaying HPV58 L2 aa.16～37 on the Surface of L1 h4-βJ coil Induces Robust Crossneutralizing Antibody Responses against HPV16,HPV52 and HPV58 in Mice

人乳头瘤病毒（Human papillomavirus,HPV）主要衣壳蛋白L1病毒样颗粒（VLP）主要诱发型别特异性中和抗体,增加L1VLP型别虽可扩大保护范围,但疫苗生产成本显著增加。HPV16,HPV52及HPV58是我国的主要高危型别。本文将HPV58次要衣壳蛋白L2aa.16-37多肽（与HPV52L2aa.16-37序列相同）插入HPV16L1的h4-βJ coil区,利用昆虫表达体系构建获得16L1h4-58dEVLP,协同人用佐剂氢氧化铝和单磷酰脂质A（Alum-MPL）免疫小鼠。活性检测结果显示16L1h4-58dE VLP免疫血清可中和17种HPV假病毒中的16种,其中针对HPV16的平均中和抗体滴度最高（滴度,76 800）,与HPV16L1VLP对照组的相当（P〉0.05）;重要的是,针对HPV58及HPV52的平均中和抗体滴度均大于103（分别为4 050和1 725）;另外,还可中等滴度中和HPV45（550）,HPV18（506）,HPV33（500）,HPV39（463）,HPV68（431）,HPV6（300）,HPV57（150）,HPV11（125）及较低滴度的中和HPV2/HPV27（40）,HPV35（38）,HPV5（25）,HPV31（13）,但对HPV59没有中和活性。因此,以本研究构建16L1h4-58dE VLP进行广谱HPV疫苗的研究,可望降低疫苗成本,具有应用前景。

Current human papillomavirus（HPV）vaccines based on the major capsid protein L1 virus-like particle（VLP）mainly induce vaccine type-specific neutralizing antibodies.Continuing to add more type of VLPs in a vaccine will raise the complexity and cost of production.High-risk HPV16,HPV58 and HPV52 are highly prevalent in China.In this study,we constructed 16 L1 h4-58 dE chimeric VLP by displaying HPV58 L2 aa.1637（100%identity with HPV52）on the h4 coil region of HPV16 L1,and assessed its immunogenicity in mice.We found that the 16 L1 h4-58 dE VLP adjuvanted with alum plus monophosphoryl lipid A could induce cross-neutralizing antibody responses against 16 out of 17 tested HPV pseudoviruses,and the titer against HPV16 was as high as that induced by HPV16 L1 VLP（titer105,P〉0.5）.More importantly,titers over 103 were observed against two high-risk HPVs including HPV58（titer,4,050）and HPV52（titer,1,725）,and medium or low titers of cross-neutralizing antibodies against other 13 tested HPV pseudoviruses,including HPV45, HPV18, HPV33, HPV39, HPV68, HPV6, HPV57,HPV11,HPV2/HPV27,HPV35,HPV5,HPV31,but not against HPV59.Our data demonstrate that the 16 L1 h4-58 dE VLP is a promising candidate for the formulation of broader-spectrum HPV vaccines.