Pluronic® F127 enhances the effect as an adjuvant of chitosan microspheres in the intranasal delivery of Bordetella bronchiseptica antigens containing dermonecrotoxin

We have studied a vaccine delivery system in vitro and in vivo based on chitosan microspheres (CMs) prepared in the presence of selected immunomodulators, Pluronic® block copolymer F127 (F127). The Bordetella bronchiseptica multiple antigens containing dermonecrotoxin (BBD), a virulent factor leading to atrophic rhinitis (AR) in swine was loaded in CMs/F127 or CMs alone. The microspheres, prepared using an ionic gelation process with tripolyphosphate, demonstrated release profiles that showed a greater amount of BBD being released from BBD-loaded CMs/F127 (BBD-CMs/F127). In vitro experiments using mouse alveolar macrophage cells (RAW 264.7) demonstrated that BBD-CMs/F127 have significantly higher immune-stimulating activities than controls. The highest immune-stimulating activities by the BBD-CMs/F127 using RAW 264.7 cells were mirrored in the in vivo studies following nasal administration to mice. The mice immunized with BBD-CMs/F127 showed higher BBD specific IgA antibody responses in nasal wash, saliva and serum than mice immunized with BBD-CMs alone. Protective immunity was measured by survival rate after challenge with B. bronchiseptica via the nasal cavity. The survival rate of the group treated with BBD-CMs/F127 was higher than those of other groups. These results suggested that CMs/F127 represents a novel mucosal delivery system and that F127 could enhance the delivery of BBD-CMs in the vaccination scheme.