Abstract
BackgroundActinobacillus pleuropneumoniae (APP) is one of the most important swine pathogens worldwide. Identification and characterization of novel antigenic APP vaccine candidates are underway. In the present study, we use an immunoproteomic approach to identify APP protein antigens that may elicit an immune response in serotype 1 naturally infected swine and serotype 1 virulent strain S259-immunized rabbits.ResultsProteins from total cell lysates of serotype 1 APP were separated by two-dimensional electrophoresis (2DE). Western blot analysis revealed 21 immunoreactive protein spots separated in the pH 4-7 range and 4 spots in the pH 7-11 range with the convalescent sera from swine; we found 5 immunoreactive protein spots that separated in the pH 4-7 range and 2 in the pH 7-11 range with hyperimmune sera from S259-immunized rabbits. The proteins included the known antigens ApxIIA, protective surface antigen D15, outer membrane proteins P5, subunit NqrA. The remaining antigens are being reported as immunoreactive proteins in APP for the first time, to our knowledge.ConclusionsWe identified a total of 42 immunoreactive proteins of the APP serotype 1 virulent strain S259 which represented 32 different proteins, including some novel immunoreactive factors which could be researched as vaccine candidates.
Highlights
Actinobacillus pleuropneumoniae (APP) is one of the most important swine pathogens worldwide
Digital images of the western blots and Ponceau Sstained membranes were compared with the 2DE gels by aligning the images with the layer function of Photoshop CS (Additional file 1, Figure S1, Additional file 2, Figure S2, Additional file 3, Figure S3, Additional file 4, Figure S4). Using this method to align developed membranes and 2DE gels in the pH 4-7 range (Figures 1, 2) and the pH 7-11 range (Figures 3, 4), we found 42 immunoreactive protein spots that represented 32 different proteins
In this study, a total of 42 immunoreactive spots, representing 32 different proteins from APP S259 were identified by western blot analysis with convalescent sera from swine and hyperimmune sera from rabbits
Summary
Actinobacillus pleuropneumoniae (APP) is one of the most important swine pathogens worldwide. Identification and characterization of novel antigenic APP vaccine candidates are underway. We use an immunoproteomic approach to identify APP protein antigens that may elicit an immune response in serotype 1 naturally infected swine and serotype 1 virulent strain S259-immunized rabbits. To control APP pathogenesis, several types of vaccines have been developed that offer various degrees of protection. Further development of cross-serovar vaccines would benefit from a molecular understanding of APP pathogenesis, which is a complex process involving a number of different potential virulence factors. The most commonly associated virulence factors [8] i.e., ApxI, ApxII, ApxIII, and ApxIV, have been tested as subunit vaccine candidates offering potential cross-serovar protection [9,10]. DNA vaccines encoding multiple Apx toxins offer a novel strategy for protecting against APP infection [11]. Lipopolysaccharide (LPS)-based vaccines are alternative to protein antigens [14]
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