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Porcine contagious pleuropneumonia (PCP) is a serious respiratory disease in the pig industry. In the present study, candidate vaccines with broad-spectrum and disease-resistant effects were screened. The App lolA gene was amplified by PCR from the Actinobacillus pleuropneumoniae (App) serotype 1 genome. The bioinformatics analysis results revealed that the App lolA consisted of 633 bp, encoding 210 amino acid proteins. In addition, the App LolA shared a high sequence identity among the different App serotypes. In this study, the App lolA was inserted into pET32a (+). The recombinant protein LolA (rLolA) was produced in E. coli BL21 and then determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and a Western Blot process. Immune protective efficacy testing of the rLolA was performed in 4-week old BALA/c mice. The serum IgG levels were assayed using an enzyme-linked immune sorbent assay (ELISA) method. The results showed that the serum IgG levels of the mice in the experimental group were significantly higher than those in the Frund incompleted adjuvant (IFA) and saline immunization groups after the third immunization. In particular, significant differences in the high dose group (50 rLolA + IFA) (P<0.01). 50 μg rLolA +IFA, 30 μg rLolA + IFA, 30 μg rLolA and 10 μg rLolA+ IFA treatments had induced 40%, 20%, 20%, 10% and 10% survival rates respectively. However, no mice had survived among the adjuvant group and saline groups. The 50 μg rLolA +IFA group displayed reductions in lung lesions. Therefore, it was ascertained from these results that the rLolA had induced partial immune protection in the experimental mice against App challenge infections. This study successfully accumulated data valuable for the future exploration of new vaccines for porcine contagious pleuropneumonia.

The immune protective efficacy of recombinant outer - membrane protein LolA from Actinobacillus pleuropneumoniae in mice