Targeting the CD40 receptor displayed by antigen-presenting cells to deliver a specific immunogen has been successfully used to enhance immune responses, specifically increasing antibody production and enhancing antibody affinity. When tested in chickens, this platform induced specific IgG and IgA production within one week post-immunization. However, proof of conferred protective efficacy using the CD40-targeting vaccination method was still undetermined. Whole avian influenza virus was loaded onto the guided complex and immunized birds were challenged with highly pathogenic avian influenza (HPAI) to test efficacy. Furthermore, this research addresses the application of guided complexes as an alternative method for epitope mapping of microbial enzymes. Short peptide segments of the Clostridium perfringens alpha toxin were loaded onto the antibody-guided complex and immunized into chickens to induce antibody production for downstream use in neutralization assays to identify specific regions able to block the toxin's enzymatic functions. Lastly, to expand the antibody-guided system repertoire, monoclonal antibodies against a new receptor, specifically dendritic cell (DC) marker CD205, were developed for potential use to further enhance immune response activation. Anti-CD205 monoclonal antibodies were used to develop a new in vitro DC system obtained from peritoneal exudate cells. In HPAI efficacy studies, functional antibody titers were detected up to six weeks after a single subcutaneous administration. When boosted, the antibody-guided complex conferred 100% protection in birds upon lethal H5N1 challenge. The guided system also proved useful for rapid polyclonal antibody production in chickens, which can be used in epitope mapping studies. This system favors linear peptide targets for immunization in order to maintain cost-effectiveness and short turnover time, but can still be used with conformational epitopes. Monoclonal antibodies were successfully constructed against chicken CD205 and used in a variety of immunoassays, as well as magnetic bead isolation of DCs from peritoneal exudate cell populations. Overall, these data are the first to report protective efficacy using the CD40-targeting system in chickens, the first to propose the use of guided complexes in epitope mapping, and the first to isolate DCs from peritoneal exudate using the anti-CD205 monoclonal antibodies.
Targeting the CD40 receptor displayed by antigen-presenting cells to deliver a specific immunogen has been successfully used to enhance immune responses, specifically increasing antibody production and enhancing antibody affinity. When tested in chickens, this platform induced specific IgG and IgA production within one week post-immunization. However, proof of conferred protective efficacy using the CD40-targeting vaccination method was still undetermined. Whole avian influenza virus was loaded onto the guided complex and immunized birds were challenged with highly pathogenic avian influenza (HPAI) to test efficacy. Furthermore, this research addresses the application of guided complexes as an alternative method for epitope mapping of microbial enzymes. Short peptide segments of the Clostridium perfringens alpha toxin were loaded onto the antibody-guided complex and immunized into chickens to induce antibody production for downstream use in neutralization assays to identify specific regions able to block the toxin's enzymatic functions. Lastly, to expand the antibody-guided system repertoire, monoclonal antibodies against a new receptor, specifically dendritic cell (DC) marker CD205, were developed for potential use to further enhance immune response activation. Anti-CD205 monoclonal antibodies were used to develop a new in vitro DC system obtained from peritoneal exudate cells.
In HPAI efficacy studies, functional antibody titers were detected up to six weeks after a single subcutaneous administration. When boosted, the antibody-guided complex conferred 100% protection in birds upon lethal H5N1 challenge. The guided system also proved useful for rapid polyclonal antibody production in chickens, which can be used in epitope mapping studies. This system favors linear peptide targets for immunization in order to maintain cost-effectiveness and short turnover time, but can still be used with conformational epitopes. Monoclonal antibodies were successfully constructed against chicken CD205 and used in a variety of immunoassays, as well as magnetic bead isolation of DCs from peritoneal exudate cell populations. Overall, these data are the first to report protective efficacy using the CD40-targeting system in chickens, the first to propose the use of guided complexes in epitope mapping, and the first to isolate DCs from peritoneal exudate using the anti-CD205 monoclonal antibodies.
