我国科学家首次解码致癌性病毒HPV58关键蛋白非构象型表位组
我国科学家首次解码致癌性病毒HPV58关键蛋白非构象型表位组
如同人类基因组解码有助于发现功能基因一样,人们也寄希望对严重威胁人类健康如甲型流感病毒和导致子宫颈癌的人乳头瘤病毒(HPV)等病毒靶蛋白的抗原表位扫描作图,以知晓它们中的全部抗体中和性表位,以用于病毒感染检测或用于研制相关疫苗。但是,尽管之前表位鉴定方法已有数百种之多,但都不能用兔和鼠类等物种多克隆抗体(多抗)鉴定每一抗体所识别的表位最小基序,因而始终无法揭示任一靶蛋白的精细表位组,导致相关研究仅能获得非常有限的结果。
由复旦大学上海市计划生育科学研究所徐万祥研究员和生命科学院谢毅教授领衔的两个团队为主,选择在国内流行仅次于致癌高风险HPV16型的HPV58型三个关键蛋白,开展了用兔多抗的表位组解码及组学研究。相关成果已于近期在《Scientific Reports》杂志上刊发。论文成果具体显示:1)使用先前自主建立的简便经济的改良生物合成肽法,解码了致癌性HPV58型三个不同长度E6、E7和L1蛋白的完整非构象型抗体表位组,第一次展现了这一研究新方向的可行性;2)在致癌性E6和E7蛋白以及预防性疫苗靶衣壳L1蛋白上发现了总共30个精细表位,包括它们在同源蛋白中的特异性和保守性,尤其发现了能覆盖绝大部分高危致癌HPV型的三个“广谱性”表位,为今后研制“通用”预防性HPV多表位肽疫苗提供了新依据;3)以往致癌性HPV型的认定或分类,都依赖妇女HPV感染和出现子宫颈癌几率的临床流行病学调查和HPV系统进化树分析结果,一直无一个客观的标准。此次在致癌性E6蛋白上发现的仅存在于高危型HPV中的一个表位,将可用作那些稀有和新发现HPV型致癌风险性判断的第一个参照标志物;4)利用鼠抗HPV58-L1假病毒颗粒样血清和生物信息学分析手段,分析了各表位的抗体可及性和潜在的抗体保护性表位。据悉,在本世纪初继基因组学和蛋白质组学之后,更提出了抗原表位组学研究新方向。但是,国内外至今未见表位组学研究的实质性启动或相关报道。此项研究为此领域提供了以往难以获得的大量有价值信息和结果,也凸显了靶蛋白非构象型抗体表位组解码和相关组学研究的意义和总体框架。研究将为真正有效的预防性重组多表位肽疫苗和诊断用抗原的研制带来新机遇。
Wan-Xiang Xu, Jian Wang, Hai-Ping Tang, Ya-Ping He, Qian-Xi Zhu, Satish K. Gupta, Shao-Hua Gu, Qiang Huang, Chao-Neng Ji, Ling-Feng Liu, Gui-Ling Li, Cong-Jian Xu
& Yi Xie. Epitomics: IgG epitome decoding of E6, E7 and L1 proteins from oncogenic human papillomavirustype 58. Sci Rep.2016 Oct 6;6:34686. doi: 10.1038/srep34686.
To enable rational multi-epitope vaccine and diagnostic antigen design, it is imperative to delineate complete IgG-epitome of the protein. Here, we describe results of IgG-epitome decoding of three proteins from high-risk (HR-) oncogenic human papillomavirus type 58 (HPV58). To reveal their entire epitomes, employing peptide biosynthetic approach, 30 precise linear B-cell epitopes (BCEs) were mapped on E6, E7 and L1 proteins using rabbits antisera to the respective recombinant proteins. Using sequence alignment based on BCE minimal motif, the specificity and conservativeness of each mapped BCE were delineated mainly among known HR-HPVs, including finding 3 broadly antibody cross-reactive BCEs of L1 that each covers almost all HR-HPVs. Western blots revealed that 13 of the 18 BCEs within L1-epitome were recognized by murine antisera to HPV58 virus-like particles, suggesting that these are antibody accessible BCEs. Also, a highly conserved epitope (YGD/XTL) of E6 was found to exist only in known common HR-HPVs, which could be used as the first peptide reference marker for judging HR-HPVs. Altogether, this study provides systemic and exhaustive information on linear BCEs of HR-HPV58 that will facilitate development of novel multi-epitope diagnostic reagents/chips for testing viral antibodies and ‘universal’ preventive HPV peptide vaccine based on L1 conserved BCEs.
来源: 中国科技网 作者: 王春（原标题：我科学家解码HPV三个编码蛋白非构象型抗体表位组）
Epitome mapping of HPV58-E6
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