title: Identification of potential CTL epitopes of bovine RSV using allele-specific peptide motifs from bovine MHC class I molecules.
abstract: Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infection in young infants and housed calves. Depletion of CD8+ lymphocytes from calves inhibited their ability to clear the virus from the nasopharynx and lungs. To study these cells further, a cytotoxic T lymphocyte (CTL) assay was established. CTL could be demonstrated in the peripheral blood of gnotobiotic calves 7-10 days post infection (p.i.) with RSV and in lungs 10 days p.i. This response was both MHC-restricted and virus-specific. Following separation of the lung lymphocytes by magnetic activated cell sorting, it was shown that the cytolytic activity was mediated by cells of the CD8+ phenotype. To identify epitopes recognised by bovine CTL, the consensus motifs from MHC class I alleles found in the herd at Compton were identified. cDNA libraries were constructed and screened for full length class I sequences. The isolated cDNA clones were then transfected into mouse P815 cells and the expressed product immunoprecipitated and matched with a serological specificity. The bovine MHC class I molecules were isolated from lysed transfected cells by affinity chromatography, using a monoclonal antibody specific for bovine MHC class I, and bound peptides were separated by reverse-phase HPLC. Analysis of the protein sequences of bovine RSV for the defined motifs has identified potential CTL epitopes.
authors.affiliation: Institute for Animal Health, Compton, Newbury, UK.
authors.forename: R M
authors.initials: RM
authors.lastname: Gaddum
authors.forename: S A
authors.initials: SA
authors.lastname: Ellis
authors.forename: A C
authors.initials: AC
authors.lastname: Willis
authors.forename: R S
authors.initials: RS
authors.lastname: Cook
authors.forename: K A
authors.initials: KA
authors.lastname: Staines
authors.forename: L H
authors.initials: LH
authors.lastname: Thomas
authors.forename: G
authors.initials: G
authors.lastname: Taylor
mesh_terms: D000483:Alleles; D000818:Animals; D002417:Cattle; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology*; D018112:Respiratory Syncytial Virus, Bovine / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000821:virology*; D014407:Tumor Cells, Cultured
chemical_list: D000939:Epitopes; D015395:Histocompatibility Antigens Class I; D010446:Peptide Fragments