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Volume 142, Issue 9

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

Abstract

Defined segments of the leukotoxin A gene () from an A1 serotype of were cloned into a plasmid vector and expressed as LacZα 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 serotype A1 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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Keyword(s): epitope , leukotoxin A , Pasteurella haemolytica and RTX toxin
© Society for General Microbiology 1996
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1996-09-01
2025-05-12
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/content/journal/micro/10.1099/00221287-142-9-2499
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Characterization of epitopes involved in the neutralization of Pasteurella haemolytica serotype A1 leukotoxin
Microbiology 142, 2499 (1996); https://doi.org/10.1099/00221287-142-9-2499
/content/journal/micro/10.1099/00221287-142-9-2499
/content/journal/micro/10.1099/00221287-142-9-2499
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