Kobasa D, Mulvey M, Lee J S, Scraba D G
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Virology. 1995 Dec 1;214(1):118-27. doi: 10.1006/viro.1995.9948.
A panel of five neutralizing monoclonal antibodies was generated from mice immunized with an attenuated strain of Mengo virus. Four of the antibodies were used to select mutants of Mengo virus which were able to escape neutralization by the selecting antibody, but it was not possible to select mutants which could escape neutralization by the fifth antibody. The capsid coding region of the RNA genome of each mutant was directly sequenced to identify the mutation(s) responsible for the neutralization escape phenotype. These results are compared to those of a previous study in which immunogenic determinants recognized by neutralizing antibodies generated against pentameric capsid subunits were located on the external surface of the Mengo virion. We have confirmed the existence of the previously identified immunogenic determinant in VP3 (site 2) as well as an immunodominant determinant in VP2 (site 1). Two previously uncharacterized determinants, located in surface loops of VP1 (sites 3 and 4A), were also identified. None of the mutations conferring the neutralization escape phenotype was found near the surface depressions on the virion which are believed to be the receptor binding sites.
用减毒的门戈病毒株免疫小鼠,产生了一组五种中和单克隆抗体。其中四种抗体用于筛选能够逃避相应筛选抗体中和作用的门戈病毒突变体,但无法筛选出能逃避第五种抗体中和作用的突变体。对每个突变体RNA基因组的衣壳编码区进行直接测序,以确定导致中和逃逸表型的突变。将这些结果与之前的一项研究结果进行比较,在之前的研究中,针对五聚体衣壳亚基产生的中和抗体识别的免疫原性决定簇位于门戈病毒粒子的外表面。我们证实了之前在VP3(位点2)中鉴定出的免疫原性决定簇以及VP2(位点1)中的一个免疫显性决定簇的存在。还鉴定出了位于VP1表面环中的两个之前未表征的决定簇(位点3和4A)。在病毒粒子上被认为是受体结合位点的表面凹陷附近,未发现赋予中和逃逸表型的突变。