Abstract
BackgroundInfection by Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae, either alone or together, causes serious respiratory diseases in pigs.ResultsTo develop an efficient multi-disease subunit vaccine against these pathogens, we produced a chimeric protein called Ap97, which comprises a deletion derivative of the N-terminal region of the A. pleuropneumoniae ApxIII toxin (ApxN) and the R1 and R2 repeats of M. hyopneumoniae P97 adhesin (P97C), using an E. coli expression system.The levels of both IgG1 and IgG2a isotypes specific for ApxN and P97C in the sera of Ap97-immunized mice increased, and Ap97 induced the secretion of IL-4 and IFN-γ by mouse splenocytes. Antisera from mice and pigs immunized with Ap97 readily reacted with both native ApxIII and P97 proteins. In addition, immunization with the Ap97 vaccine effectively protected pigs against challenge with both pathogens.ConclusionsThese findings suggest that Ap97 confers immunogenicity, and is an effective vaccine that protects pigs against infection by M. hyopneumoniae and A. pleuropneumoniae.
Highlights
Infection by Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae, either alone or together, causes serious respiratory diseases in pigs
Production of recombinant proteins SDS-PAGE analysis of proteins purified after expression in E. coli using pET-ApxIII toxin (ApxN), pET-P97C, and pET-Ap97 vectors revealed prominent bands at approximately 15 kDa, 45 kDa, and 60 kDa, respectively (Figure 2A)
Antiserum against the Ap97 vaccine reacted well with ApxN, P97C, and Ap97, demonstrating that immunization with Ap97 induces the production of antibodies that bind to all three recombinant proteins
Summary
Infection by Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae, either alone or together, causes serious respiratory diseases in pigs. Based on differences in the capsular polysaccharides, 15 serotypes of A. pleuropneumoniae have been identified to date, which show significant variation with respect to virulence and geographical distribution. Most of those serotypes secrete one or more A. pleuropneumoniae toxin (Apx), identified as ApxI, ApxII, ApxIII, and ApxIV. A. pleuropneumoniae vaccines include inactivated wholecell bacterins as well as the more promising subunit vaccines and live attenuated vaccines [4]. Most commercially available vaccines against A. pleuropneumoniae infection comprise inactivated whole-cell bacterins derived from various serotypes of A. pleuropneumoniae [5]. Live attenuated vaccines present a number of bio-safety concerns
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