HPV18 L2保守中和表位肽协同MF59与CpG-ODN免疫可诱发广谱中和抗体反应  

Conservative Neutralizing Epitope of Human Papillomavirus Type 18 L2 Immunized with MF59 and CpG-ODN Can Induce Broad-spectrum Neutralizing Antibodies

作　　者：马铭饶 郝亚茹 张婷[1] 刘洪洋[1] 王志荣 夏百成 周艳[1] 许雪梅[1] MA Ming-rao;HAO Ya-ru;ZHANG Ting;LIU Hong-yang;WANG Zhi-rong;XIA Bai-cheng;ZHOU Yan;XU Xue-mei(Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences,Pecking Union Medical College,Beijing,100005,China)

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院,北京100005

出　　处：《现代生物医学进展》2020年第20期3801-3806,共6页Progress in Modern Biomedicine

基　　金：中国医学科学院医学与健康科技创新工程项目基金(2016-I2M-3-026);中国医学科学院医学与健康科技创新工程项目基金(2020-I2M-2-014);国家自然科学基金面上项目(31970867)。

摘　　要：目的:人乳头瘤病毒(HPV)次要外壳蛋白L2保守中和表位肽可诱发交叉中和抗体,研究L2保守中和表位肽免疫原性的特点利于HPV通用疫苗的研发。HPV18是第二常见的优势流行高危型,但18L2保守表位肽的免疫活性未见报道。方法:本研究采用化学法合成HPV18 L2N端多肽(18RG-1)并偶联KLH获得18RG1-KLH肽;联合MF59/CpG-ODN复合佐剂或弗氏佐剂皮下免疫BALB/c小鼠5次,用假病毒中和实验检测抗血清针对α6、α7、α9及α11亚属中多个致癌型HPV的中和抗体。结果:MF59/CpG-ODN复合佐剂多肽组抗血清对所有6种检测型别的中和活性与弗氏佐剂多肽组的相当。MF59/CpG-ODN佐剂多肽组抗血清具有广谱中和活性,中和范围至少包括14种致癌型HPV,中和抗体滴度最高的为HPV45(438)和HPV18(325),其次为HPV68(163)和HPV70(150),这四种优势中和型别均为α7亚属。结论:首次发现HPV18 L2 RG1保守中和表位免疫血清可诱发广谱中和抗体反应(其中对α7亚属的HPV中和活性最强,为优势中和型别)。研究结果为基于L2保守表位的广谱HPV疫苗研发奠定基础。Objective: The human papillomavirus(HPV) minor coat protein L2 contains conservative neutralizing epitope which can induce cross-neutralizing antibodies. It is beneficial to analyze the immunogenicity of L2 conservative neutralizing epitopes for the development of HPV universal vaccines. HPV18 is the second most prevalent high-risk type worldwide, but the immunogenicity of the 18 L2 peptide has not been reported. Methods: HPV18 L2 peptide(18 RG-1) was synthesized by chemical methods and conjugated with KLH to construct 18 RG1-KLH;BALB/c mice were immunized five times with MF59/CpG-ODN or Freud’s Adjuvant. Neutralizing antibodies against oncogenic HPVs from α6, α7, α9 and α11 species were detected by pseudovirus neutralization assay. Results: The neutralizing activity of the antiserum of the MF59/CpG-ODN group against all 6 detected types is equivalent to that of the Freund’s Adjuvant group. The antiserum of the MF59/CpG-ODN group,which can at least neutralize 14 oncogenic HPVs, has broad-spectrum neutralizing activity. The highest neutralizing antibody titers are against HPV45(438) and HPV18(325), followed by HPV68(163) and HPV70(150), all belong to α7 specie. Conclusions: It is discovered for the first time that conservative neutralizing epitope HPV18 L2 RG1 can induce broad-spectrum neutralizing activity(neutralizing activity against α7 species is the strongest). The results lay the foundation of the development of broad-spectrum HPV vaccines based on L2 conserved epitopes.

关 键 词：人乳头瘤病毒18型 L2保守中和表位 广谱中和抗体 

分 类 号：R-33[医药卫生] R730.261