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Identification of Foot-and-Mouth Disease Virus-Specific Linear B-Cell Epitopes To Differentiate between Infected and Vaccinated Cattle

Bettina-Judith Höhlich,^1, Karl-Heinz Wiesmüller,² Tobias Schlapp,³ Bernd Haas,¹ Eberhard Pfaff,¹ and Armin Saalmüller^1*

Institut für Immunologie, Bundesforschungsanstalt für Viruskrankheiten der Tiere, 72076 Tübingen,¹ EMC Microcollections, 72070 Tübingen,² Bayer Animal Health, D-51368 Leverkusen, Germany³

Received 10 February 2003/ Accepted 21 May 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 O₁K in a peptide enzyme-linked immunosorbent assay, allowed the identification of 12 FMDV strain O₁K-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.

Present address: Miltenyi Biotec, 51429 Bergisch Gladbach, Germany.

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