Expression Analysis of Outer Membrane Protein HPS_06257 in Different Strains of Glaesserella parasuis and Its Potential Role in Protective Immune Response against HPS_06257-Expressing Strains via Antibody-Dependent Phagocytosis

SIMPLE SUMMARY: Glaesserella parasuis, one of the opportunistic pathogens causing Glässer’s disease in piglets, has become a significant concern for pig farmers. Vaccination has been shown to be effective in preventing Glaesserella parasuis infection by inducing the protective immune response. Notably, a humoral immune response plays an important role in protection of Glaesserella parasuis infection. The mechanism of protection by antibodies has been shown to be associated with antibody-opsonized phagocytosis, which facilitates uptake of Glaesserella parasuis by phagocytes such as macrophages. Outer membrane proteins of Glaesserella parasuis, as the promising candidates, are often chosen to develop subunit vaccines. HPS_06257 is one of the outer membrane proteins that has been shown to confer protection against Glaesserella parasuis infection. However, little is known about the role of HPS_06257 in the protective immune response. We demonstrate that antibody-dependent phagocytosis is involved in the protective effects of HPS_06257. Our findings extend our understanding of how antibody-dependent phagocytosis may contribute to the immune protection afforded by other outer membrane proteins. Thus, our study provides insight into the protective antigens of Glaesserella parasuis and useful information for the development of novel vaccines to prevent Glaesserella parasuis infection. ABSTRACT: HPS_06257 has been identified as an important protective antigen against Glaesserella parasuis infection. However, little is known about the role of HPS_06257 in the protective immune response. A whole-genome data analysis showed that among 18 isolates of Glaesserella parasuis, 11 were positive for the HPS_06257 gene, suggesting that not every strain contains this gene. We used PCR to investigate the presence of the HPS_06257 gene among 13 reference strains and demonstrated that 5 strains contained the gene. A polyclonal antibody against HPS_06257 was generated with a recombinant protein to study the expression of HPS_06257 in those 13 strains. Consistent with the PCR data, five strains expressed HPS_06257, whereas eight strains were HPS_06257 null. We also compared the protective effects of HPS_06257 against an HPS_06257-expressing strain (HPS5) and an HPS_06257-null strain (HPS11). Immunization with HPS_06257 only protected against HPS5 and not HPS11. Moreover, phagocytosis of antibody-opsonized bacteria demonstrates that the antibody against HPS_06257 increased the phagocytosis of the HPS5 strain by macrophages but not the phagocytosis of the HPS11 strain, suggesting that antibody-dependent phagocytosis is responsible for the protective role exerted by HPS_06257 in the immune response to HPS5. Our data also show that the antibody against HPS_06257 increased the phagocytosis of the other HPS_06257-expressing strains by macrophages but not that of HPS_06257-null strains. In summary, our findings demonstrate that antibody-dependent phagocytosis contributes to the protective immune response induced by immunization with HPS_06257 against HPS_06257-expressing strains.