Abstract
Leukotoxin produced by Mannheimia (Pasteurella) haemolytica is an important virulence factor in shipping fever pneumonia in feedlot cattle and is a critical protective antigen. In this study, the immune response to a chimeric protein generated by combining a gene fragment encoding neutralizing epitopes of M. haemolytica leukotoxin and a fimbrial protein gene (fim N) from Bordetella bronchiseptica was evaluated. The recombinant gene was cloned in a bacterial expression vector under the control of the tac promoter and expressed as a fusion protein with glutathione-S-transferase (GST) in Escherichia coli. Immunization of mice with the recombinant protein, GST-LTXFIM elicited a significantly stronger anti-leukotoxin antibody response than comparable immunizations with GST-LTX fusion proteins lacking FIM N. The GST-LTXFIM was also more stable than GST-LTX during storage at -80 degrees C, thus alleviating a stability problem inherent to leukotoxin. This chimeric protein may be a candidate for inclusion in new generation vaccines against shipping fever pneumonia.
Publication types
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Comparative Study
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Evaluation Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Animals
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Antibodies, Bacterial / biosynthesis
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Antibodies, Bacterial / immunology
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Antibody Specificity
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Antigens, Bacterial / genetics
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Antigens, Bacterial / immunology*
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Bacterial Proteins*
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Bacterial Vaccines / immunology*
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Bordetella bronchiseptica / immunology*
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Cattle
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Cattle Diseases / prevention & control
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Epitopes / immunology
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Exotoxins / genetics
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Exotoxins / immunology*
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Female
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Fimbriae Proteins*
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Fimbriae, Bacterial / immunology*
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Genes, Synthetic
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Genetic Vectors / genetics
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Glutathione Transferase / genetics
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Hemolysin Proteins / genetics
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Hemolysin Proteins / immunology*
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Mannheimia haemolytica / genetics
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Mannheimia haemolytica / immunology*
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Mannheimia haemolytica / pathogenicity
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Mice
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Mice, Inbred BALB C
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Neutralization Tests
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Pasteurellosis, Pneumonic / prevention & control
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Rabbits
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / isolation & purification
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Species Specificity
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Vaccines, Synthetic
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Virulence
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Virulence Factors, Bordetella*
Substances
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Antibodies, Bacterial
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Antigens, Bacterial
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Bacterial Proteins
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Bacterial Vaccines
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Epitopes
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Exotoxins
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FimN protein, Bordetella bronchiseptica
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Hemolysin Proteins
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Recombinant Fusion Proteins
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Vaccines, Synthetic
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Virulence Factors, Bordetella
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lktA protein, bacteria
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Fimbriae Proteins
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Glutathione Transferase