Identification of T-cell epitopes in the structural and non-structural proteins of classical swine fever virus.
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-gamma 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.
Armengol
Elisenda
E
Institute of Immunology, Federal Research Centre for Virus Diseases of Animals, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Germany1.
Wiesmüller
Karl-Heinz
KH
EMC microcollections GmbH, D-72070 Tübingen, Germany2.
Wienhold
Daniel
D
Institute of Immunology, Federal Research Centre for Virus Diseases of Animals, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Germany1.
Büttner
Mathias
M
Institute of Immunology, Federal Research Centre for Virus Diseases of Animals, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Germany1.
Pfaff
Eberhard
E
Institute of Immunology, Federal Research Centre for Virus Diseases of Animals, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Germany1.
Jung
Günther
G
Institut für Organische Chemie, Universität Tübingen, D-72076 Tübingen, Germany3.
Saalmüller
Armin
A
Institute of Immunology, Federal Research Centre for Virus Diseases of Animals, Paul-Ehrlich-Straße 28, D-72076 Tübingen, Germany1.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry* / Q000276:immunology*; D002455:Cell Division; D002478:Cells, Cultured; D006692:Classical Swine Fever Virus / Q000737:chemistry* / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D016679:Genome, Viral; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D013552:Swine / Q000276:immunology* / Q000821:virology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology; D015678:Viral Structural Proteins / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D010455:Peptides; D014764:Viral Proteins; D015678:Viral Structural Proteins; D007371:Interferon-gamma