THE OPEN READING FRAME L2 OF COTTONTAIL RABBIT PAPILLOMAVIRUS CONTAINS ANTIBODY-INDUCING NEUTRALIZING EPITOPES
THE OPEN READING FRAME L2 OF COTTONTAIL RABBIT PAPILLOMAVIRUS CONTAINS ANTIBODY-INDUCING NEUTRALIZING EPITOPES
复制标题
DOI：
10.1016/0042-6822(91)90890-n
复制
发表时间：
1991-04-01
期刊：
影响因子：
3.7
通讯作者：
DIANGELO, SL
中科院分区：
文献类型：
--
作者：
CHRISTENSEN, ND;KREIDER, JW;DIANGELO, SL
用棉尾兔乳头瘤病毒(CRPV)衣壳蛋白的开放阅读框(ORF)融合蛋白制备多克隆抗血清。将CRPVL1的羧基末端2/3和CRPVL2的羧基末端的一半克隆到细菌表达载体中，以诱导蛋白作为抗原和免疫原。多克隆抗血清进行了一系列的免疫学检测，包括ELISA法、Western印迹法和CRPV中和试验。ELISA法表明，表达的L1蛋白多克隆抗血清与破碎型CRPV病毒粒子有较强的反应，而与完整的CRPV病毒粒子和破碎型BPV-1病毒粒子反应较弱。抗CRPV L2抗血清仅与完整和破损的CRPV病毒粒子抗原发生强烈反应。用这些多克隆抗血清在HPV-11、BPV-1和CRPV病毒颗粒的Western blotting中检测到CRPV的衣壳蛋白。抗L1血清识别60 kDa的主要衣壳蛋白，抗L2血清识别76 kDa的CRPV病毒蛋白。只有针对表达的L2产生的抗血清中和CRPV。然而，抗L2血清的中和滴度比用完整的CRPV病毒粒子免疫产生的中和多克隆抗血清低几个数量级。
Polyclonal antisera were generated against bacterially derived fusion proteins of the open reading frames (ORFs) of the capsid proteins of cottontail rabbit papillomavirus (CRPV). The carboxy-terminal two-thirds of CRPV L1 and the carboxy-terminal half of CRPV L2 were cloned into a bacterial expression vector and induced proteins were used as antigen and immunogen. The polyclonal antisera were tested in a series of immunological assays, including ELISA, Western blot, and neutralization of CRPV. ELISA demonstrated that the polyclonal antisera raised against expressed L1 proteins reacted strongly to disrupted CRPV virion antigen and weakly both to intact CRPV virion and disrupted BPV-1 virion. Anti-CRPV L2 antisera reacted strongly only to intact and disrupted CRPV virion antigen. Viral capsid proteins of CRPV were detected in Western blots of HPV-11, BPV-1, and CRPV virus particles by these polyclonal antisera. The anti-L1 sera recognized the major capsid protein (60 kDa) and the anti-L2 sera identified a 76-kDa viral protein of CRPV. Only the antisera generated against expressed L2 neutralized CRPV. The neutralizing titer of the anti-L2 sera, however, was several orders of magnitude lower than the titer of a neutralizing polyclonal antiserum that was generated by immunizations with intact CRPV virions.