Balasuriya U B, Rossitto P V, DeMaula C D, MacLachlan N J
Department of Veterinary Pathology, School of Veterinary Medicine, University of California, Davis 95616.
J Gen Virol. 1993 Nov;74 ( Pt 11):2525-9. doi: 10.1099/0022-1317-74-11-2525.
A panel of six neutralizing murine monoclonal antibodies (MAbs) to equine arteritis virus (EAV) was produced. The MAbs were characterized by Western immunoblotting assay and competitive ELISA. The six MAbs identify a single neutralization site on a 29K envelope glycoprotein. Deglycosylation of viral proteins prior to immunoblotting showed that the 29K protein is the glycosylated form of a 20K protein. Equine anti-EAV serum also strongly bound the 29K glycoprotein, as well as an unglycosylated protein of 17K. The equine antisera to EAV blocked the binding of a selected MAb to EAV, whereas normal equine serum did not. Two neutralization-resistant escape mutant (EM) variants of the EAV prototype were produced using MAb 6D10. The phenotypic properties of the EM viruses were characterized by neutralization and immunoblotting assays with two MAbs (6D10 and 5G11). The two MAbs failed to neutralize either EM virus, and they did not react in an immunoblot assay with any proteins of the EM viruses. In contrast, binding of the equine antiserum to viral proteins was equivalent with prototype and EM virus strains. These data clearly indicate that a 29K envelope glycoprotein expresses at least one neutralization determinant of EAV.
制备了一组针对马动脉炎病毒（EAV）的六种中和性鼠单克隆抗体（MAb）。通过Western免疫印迹分析和竞争性ELISA对这些单克隆抗体进行了表征。这六种单克隆抗体识别29K包膜糖蛋白上的一个单一中和位点。免疫印迹前对病毒蛋白进行去糖基化显示，29K蛋白是20K蛋白的糖基化形式。马抗EAV血清也强烈结合29K糖蛋白以及17K的非糖基化蛋白。马抗EAV血清可阻断所选单克隆抗体与EAV的结合，而正常马血清则不能。使用单克隆抗体6D10产生了EAV原型的两个抗中和逃逸突变体（EM）变体。通过用两种单克隆抗体（6D10和5G11）进行中和和免疫印迹分析来表征EM病毒的表型特性。这两种单克隆抗体均不能中和任何一种EM病毒，并且它们在免疫印迹分析中不与EM病毒的任何蛋白质发生反应。相比之下，马抗血清与病毒蛋白的结合在原型病毒株和EM病毒株中是相同的。这些数据清楚地表明，29K包膜糖蛋白表达EAV的至少一个中和决定簇。
