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Identification of T-cell epitopes in the structural and non-structural proteins of classical swine fever virus

Institute of Immunology, Federal Research Centre for Virus Diseases of Animals, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Germany¹
EMC microcollections GmbH, D-72070 Tübingen, Germany²
Institut für Organische Chemie, Universität Tübingen, D-72076 Tübingen, Germany³

Author for correspondence: Armin Saalmüller. Fax +49 7071 967303. e-mail armin.saalmueller{at}tue.bfav.de

To identify new T-cell epitopes of classical swine fever virus (CSFV), 573 overlapping, synthetic pentadecapeptides spanning 82% of the CSFV (strain Glentorf) genome sequence were synthesized and screened. In proliferation assays, 26 peptides distributed throughout the CSFV viral protein sequences were able to induce specific T-cell responses in PBMCs from a CSFV-Glentorf-infected d/d haplotype pig. Of these 26 peptides, 18 were also recognized by PBMCs from a CSFV-Alfort/187-infected d/d haplotype pig. In further experiments, it could be shown that peptide 290 (KHKVRNEVMVHWFDD), which corresponds to amino acid residues 1446–1460 of the CSFV non-structural protein NS2–3 could induce interferon- secretion after secondary in vitro restimulation. The major histocompatibility complex (MHC) restriction for stimulation of T-cells by this pentadecapeptide was identified as being mainly MHC class II and partially MHC class I. In cytolytic assays, CSFV-specific cytotoxic T-lymphocytes (CTLs) were able to lyse peptide 290-loaded target cells. These findings indicate the existence of a CSFV-specific helper T-cell epitope and a CTL epitope in this peptide.

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