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Characterization of epitopes involved in the neutralization of Pasteurella haemolytica serotype A1 leukotoxin



Characterization of epitopes involved in the neutralization of Pasteurella haemolytica serotype A1 leukotoxin



Microbiology 142: 2499-2507



Defined segments of the leukotoxin A gene (IktA) from an A1 serotype of Pasteurella haemolytica were cloned into a plasmid vector and expressed as LacZ-alpha fusion proteins. These fusion proteins were electrophoresed in SDS-PAGE gels and their immunoblotting reactivities with several monoclonal antibodies characterized. The epitope recognized by a strongly neutralizing monoclonal antibody was localized to a 32 amino acid region near the C terminus of the leukotoxin A (LktA) molecule. The epitope recognized by a non-neutralizing antibody was localized to a 33 amino acid region immediately adjacent. Smaller recombinant peptides containing these epitopes were not antigenic, but a polypeptide encompassing 229 amino acids at the C terminus evoked neutralizing antibodies when used to immunize specific-pathogen-free lambs. The distributions of linear epitopes recognized by this antiserum and by antisera raised to full-length recombinant LktA and to native LktA produced by P. haemolytica serotype Al were determined by their reactivities with a set of overlapping 10 amino acid synthetic peptides. This revealed a complex distribution of linear epitopes at the C-terminal end of LktA. Toxin-neutralizing antibodies in convalescent sheep serum were shown to be directed against conformational epitopes by selective absorption of antibodies directed against linear epitopes.

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Accession: 002775626

Download citation: RISBibTeXText

PMID: 8828217

DOI: 10.1099/00221287-142-9-2499


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