Immunization of mice with delayed lysis Salmonella expressing PCV2b Cap protein enhanced mucosal and innate immunity and reduced viral load

Abstract

Porcine circovirus type 2 (PCV2) has been recognized as a critical pathogen associated with numerous porcine circovirus-related diseases. Immunization is commonly regarded as the most efficient method to combat PCV2 infection. The virus's main antigen is the Cap protein, which is encoded by the ORF2 gene. In this investigation, we cloned the PCV2 Cap gene into the pYA3681 plasmid and subsequently electrotransferred it into the delayed lysis Salmonella strain χ11802, ultimately generating the χ11802 (pYA3681-Cap) vaccine candidate strain. We assessed the levels of sIgA and IgG specific to the Cap protein in mice, revealing that their mucosal and humoral immunity had been activated by χ11802 (pYA3681-Cap). The result was a substantial elevation in antibody levels, and a notable reduction in viral load in the immunized group compared to the unvaccinated group. Furthermore, the study revealed a significant decrease in the viral load in the lungs, liver, and spleen of mice inoculated with χ11802 (pYA3681-Cap), in comparison to both the empty carrier group and the phosphate buffered saline control group. This study further investigated the lytic effect of the delayed lysis vaccine vector. The χ11802 (pYA3681-EGFP) strain was undetectable 10 days post-challenge, indicating that the vaccine strain can effectively release the carried exogenous antigen and prevent the residual vaccine strain from spreading and causing pollution in the environment. A successful construction of the χ11802 (pYA3681-Cap) strain expressing the PCV2 Cap protein was executed in this study. To summarize, our study suggests that PCV2-Cap, when expressed in the delayed lysis Salmonella strain χ11802, could serve as a safe and economically efficient candidate PCV2 vaccine.