Molecular characterization of a common 48-kilodalton outer membrane protein of Actinobacillus pleuropneumoniae

Abstract

Previous studies have shown that a vaccine prepared from outer membranes of Actinobacillus pleuropneumoniae serotype 5 can elicit protective immunity in swine against challenge with either serotype 5 or serotype 1. These results suggest the presence of common subcapsular surface antigens, such as outer membrane proteins, that contribute to cross-protective immunity. We have identified a 48-kDa outer membrane protein that is common to all 12 capsular serotypes of A. pleuropneumoniae but is not present in the outer membranes of related species of gram-negative swine pathogens. This protein is immunogenic in swine infected with either A. pleuropneumoniae serotype 5 or 1A, as well as in swine vaccinated with A. pleuropneumoniae serotype 5 outer membranes. This 48-kDa protein is readily detected in outer membranes produced by sucrose density gradient centrifugation, but it is sarcosyl soluble and therefore not found in outer membranes prepared by detergent treatment. The gene encoding the 48-kDa outer membrane protein has been cloned from A. pleuropneumoniae serotype 5 and and has been designated aopA, for Actinobacillus outer membrane protein A. The gene is 1,347 bp in length and encodes a protein, designated AopA, of 449 amino acids with a predicted molecular weight of 48,603. Southern blot analysis under high-stringency conditions showed that strains of all 12 serotypes of A. pleuropneumoniae contain DNA homologous to this gene, as do strains of two closely related species, Actinobacillus suis and Pasteurella multocida. Whether antibodies against the AopA antigen contribute to cross-protective immunity against A. pleuropneumoniae infection remains to be determined.