Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / immunology
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Antigens, Viral / chemistry*
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Antigens, Viral / immunology*
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Cell Division
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Cells, Cultured
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Classical Swine Fever Virus / chemistry*
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Classical Swine Fever Virus / immunology*
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Epitopes, T-Lymphocyte / chemistry
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Epitopes, T-Lymphocyte / immunology*
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Genome, Viral
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Histocompatibility Antigens Class I / immunology
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Histocompatibility Antigens Class II / immunology
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Interferon-gamma / immunology
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Interferon-gamma / metabolism
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Lymphocyte Activation
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Mice
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Molecular Sequence Data
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Peptides / chemical synthesis
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Peptides / chemistry
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Peptides / immunology
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Swine / immunology*
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Swine / virology*
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T-Lymphocytes, Cytotoxic / immunology
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T-Lymphocytes, Cytotoxic / metabolism
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T-Lymphocytes, Helper-Inducer / immunology
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T-Lymphocytes, Helper-Inducer / metabolism
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Viral Proteins / chemistry
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Viral Proteins / immunology
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Viral Structural Proteins / chemistry
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Viral Structural Proteins / immunology
Substances
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Antibodies, Monoclonal
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Antigens, Viral
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Epitopes, T-Lymphocyte
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Histocompatibility Antigens Class I
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Histocompatibility Antigens Class II
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Peptides
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Viral Proteins
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Viral Structural Proteins
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Interferon-gamma