Abstract
Actinobacillus pleuropneumoniae is an important veterinary pathogen that causes porcine pleuropneumonia. Lipoproteins of bacterial pathogens play pleiotropic roles in the infection process. In addition, many bacterial lipoproteins are antigenic and immunoprotective. Therefore, characterization of lipoproteins is a promising strategy for identification of novel vaccine candidates or diagnostic markers. We cloned 58 lipoproteins from A. pleuropneumoniae JL03 (serovar 3) and expressed them in Escherichia coli. Five proteins with strong positive signals in western blotting analysis were used to immunize mice. These proteins elicited significant antibody responses, and three of them (APJL_0922, APJL_1380 and APJL_1976) generated efficient immunoprotection in mice against lethal heterologous challenge with A. pleuropneumoniae 4074 (serovar 1), both in the active and passive immunization assays. Then immunogenicity of these three lipoproteins (APJL_0922, APJL_1380 and APJL_1976) were further tested in pigs. Results showed that these proteins elicited considerable humoral immune responses and effective protective immunity against virulent A. pleuropneumoniae challenge. Our findings suggest that these three novel lipoproteins could be potential subunit vaccine candidates.
Highlights
Actinobacillus pleuropneumoniae is an important veterinary pathogen that causes porcine pleuropneumonia
Based on our previous bioinformatics prediction, 60 lipoproteins were identified from the A. pleuropneumoniae JL03 (Table 1) genome for further investigation
Since lipoproteins Lip[40] and PalA have been analyzed before[18,19], here, the remaining 58 open reading frames were amplified by polymerase chain reaction (PCR) (Table S1) and E. coli expression vectors were constructed, to generate recombinant proteins
Summary
Actinobacillus pleuropneumoniae is an important veterinary pathogen that causes porcine pleuropneumonia. Five proteins with strong positive signals in western blotting analysis were used to immunize mice These proteins elicited significant antibody responses, and three of them (APJL_0922, APJL_1380 and APJL_1976) generated efficient immunoprotection in mice against lethal heterologous challenge with A. pleuropneumoniae 4074 (serovar 1), both in the active and passive immunization assays. Immunogenicity of these three lipoproteins (APJL_0922, APJL_1380 and APJL_1976) were further tested in pigs. Results showed that these proteins elicited considerable humoral immune responses and effective protective immunity against virulent A. pleuropneumoniae challenge. PGEX-KG carrying the coding sequence for lipoprotein of A. pleuropneumoniae JL03, for the expression of GST-fusion recombinant protein
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