Höhlich Bettina-Judith, Wiesmüller Karl-Heinz, Schlapp Tobias, Haas Bernd, Pfaff Eberhard, Saalmüller Armin
Institut für Immunologie, Bundesforschungsanstalt für Viruskrankheiten der Tiere, 72076 Tübingen, Germany.
J Virol. 2003 Aug;77(16):8633-9. doi: 10.1128/jvi.77.16.8633-8639.2003.
Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals. For several years, vaccination of animals, which had proven to be successful for the eradication of the disease, has been forbidden in the United States and the European Community because of the difficulty of differentiating between vaccinated and infected animals. In this study, detailed investigations of the bovine humoral immune response against FMD virus (FMDV) were performed with the aim of identifying viral epitopes recognized specifically by sera derived from FMDV-infected animals. The use of overlapping 15-mer synthetic peptides, covering the whole open reading frame of FMDV strain O(1)K in a peptide enzyme-linked immunosorbent assay, allowed the identification of 12 FMDV strain O(1)K-specific linear B-cell epitopes. Six of these linear B-cell epitopes, located in the nonstructural proteins, were used in further assays to compare the reactivities of sera from vaccinated and infected cattle. Antibodies recognizing these peptides could be detected only in sera derived from infected cattle. In further experiments, the reactivity of the six peptides with sera from animals infected with different strains of FMDV was tested, and strain-independent infection-specific epitopes were identified. Thus, these results clearly demonstrate the ability of a simple peptide-based assay to discriminate between infected and conventionally FMD-vaccinated animals.
口蹄疫(FMD)是偶蹄动物的一种高度传染性病毒性疾病。多年来,由于难以区分接种疫苗的动物和感染动物,在美国和欧洲共同体,已证明对根除该疾病有效的动物疫苗接种被禁止。在本研究中,对牛针对口蹄疫病毒(FMDV)的体液免疫反应进行了详细研究,目的是鉴定FMDV感染动物血清特异性识别的病毒表位。在肽酶联免疫吸附试验中使用覆盖FMDV O(1)K株整个开放阅读框的重叠15聚体合成肽,鉴定出12个FMDV O(1)K株特异性线性B细胞表位。其中位于非结构蛋白中的6个线性B细胞表位用于进一步试验,以比较接种疫苗和感染牛血清的反应性。仅在感染牛的血清中能检测到识别这些肽的抗体。在进一步实验中,测试了这6种肽与感染不同FMDV毒株动物血清的反应性,鉴定出与毒株无关的感染特异性表位。因此,这些结果清楚地证明了一种基于简单肽的检测方法能够区分感染动物和传统接种FMD疫苗的动物。