Pfaff E, Thiel H J, Beck E, Strohmaier K, Schaller H
Microbiology and Zentrum für Molekulare Biologie Heidelberg, University of Heidelberg, Federal Republic of Germany.
J Virol. 1988 Jun;62(6):2033-40. doi: 10.1128/JVI.62.6.2033-2040.1988.
For the investigation of the antigenic determinant structure of foot-and-mouth disease virus (FMDV), neutralizing monoclonal antibodies (MAbs) against complete virus were characterized by Western blot (immunoblot), enzyme immunoassay, and competition experiments with a synthetic peptide, isolated coat protein VP1, and viral particles as antigens. Two of the four MAbs reacted with each of these antigens, while the other two MAbs recognized only complete viral particles and reacted only very poorly with the peptide. The four MAbs showed different neutralization patterns with a panel of 11 different FMDV strains. cDNA-derived VP1 protein sequences of the different strains were compared to find correlations between the primary structure of the protein and the ability of virus to be neutralized. Based on this analysis, it appears that the first two MAbs recognized overlapping sequential epitopes in the known antigenic site represented by the peptide, whereas the two other MAbs recognized conformational epitopes. These conclusions were supported and extended by structural analyses of FMDV mutants resistant to neutralization by an MAb specific for a conformational epitope. These results demonstrate that no amino acid exchanges had occurred in the primary antigenic site of VP1 but instead in the other coat proteins VP2 and VP3, which by themselves do not induce neutralizing antibodies.
为研究口蹄疫病毒(FMDV)的抗原决定簇结构,通过蛋白质免疫印迹法(免疫印迹)、酶免疫测定以及与合成肽、分离的衣壳蛋白VP1和病毒颗粒作为抗原的竞争实验,对口蹄疫病毒完整病毒的中和单克隆抗体(MAb)进行了表征。四种单克隆抗体中的两种与这些抗原中的每一种都发生反应,而另外两种单克隆抗体仅识别完整的病毒颗粒,并且与该肽的反应非常弱。这四种单克隆抗体对一组11种不同的口蹄疫病毒株表现出不同的中和模式。比较了不同毒株的cDNA衍生的VP1蛋白序列,以寻找蛋白质一级结构与病毒中和能力之间的相关性。基于该分析,似乎前两种单克隆抗体识别由该肽代表的已知抗原位点中的重叠连续表位,而另外两种单克隆抗体识别构象表位。这些结论得到了对构象表位特异性单克隆抗体中和抗性的FMDV突变体进行结构分析的支持和扩展。这些结果表明,VP1的主要抗原位点未发生氨基酸交换,而是发生在其他衣壳蛋白VP2和VP3中,它们自身不会诱导中和抗体。