Aluminum Oxide Nanoparticle-Adjuvanted Pasteurella multocida B:2 Vaccine is Potent and Efficacious and is Able with Keyhole Limpet Hemocyanin as a Marker to Differentiate Infected from Vaccinated Animals (DIVA)

Abstract

Pasteurella multocida B:2 causes hemorrhagic septicemia (HS) in bovines in Asia. For effective implementation of HS control program in India, a safe and efficacious vaccine to provide long lasting protection and allow differentiation of infected from vaccinated animals (DIVA) has to be developed. The objectives of the present study were to develop HS-DIVA vaccine with a companion immunodiagnostic test using keyhole limpet hemocyanin (KLH) as a marker, and to improve its potency and efficacy using aluminum oxide nanoparticles (Al 2 O 3 NPs) as adjuvant in a mouse model. The experimental vaccine was constructed by mixing Al 2 O 3 NPs and KLH with formalin-inactivated P. multocida B:2 P52 strain. Swiss albino mice (n = 8/group) were inoculated twice at three-week interval. Sixteen days later, the mice were challenged with 50 × lethal dose 50 of virulent P. multocida B:2 P52 strain and protection levels were determined for each group. Antibody levels against bacterial antigens and KLH were determined by indirect ELISA. The mice in Al 2 O 3 NPs + KLH + bacterin [HSVac-I] group showed 100% protection. While, Al 2 O 3 NPs + bacterin [HSVac-II], KLH + bacterin [HSVac-III], bacterin [HSVac-IV] and alum-precipitated bacterin [HSVac-V] groups had 87.5, 62.5, 62.5 and 12.5% protection, respectively. Anti-bacterial antibody levels in the HSVac -I, -II and -III groups were significantly higher than those in HSVac-IV (*P < 0.05) and HSVac-V (**P < 0.001) groups on 14, 21, 28 and 35 days post-immunization. Anti-KLH antibodies had high titres in all the KLH-positive groups and persisted as long as antibacterial antibodies. In conclusion, KLH was suitable as a marker for HS-DIVA vaccine and with Al 2 O 3 NPs improved the HS vaccine potency and efficacy. Keywords: Al 2 O 3 nanoparticles, KLH marker antigen, bovine hemorrhagic septicemia, DIVA vaccine, Pasteurella multocida B:2 Cite this Article Ray Susim Mukul, Singh Ajit . Aluminum oxide nanoparticle-adjuvanted Pasteurella Multocida B:2 vaccine is potent and efficacious and is able with keyhole limpet hemocyanin as a marker to differentiate infected from vaccinated animals (DIVA). Research and Reviews: Journal of Immunology (RRJoI). 2015; 5(2): 8–16p.