Chitosan/sulfated locust bean gum nanoparticles: In vitro and in vivo evaluation towards an application in oral immunization.

This work proposes the design of nanoparticles based on locus bean gum (LBG) and chitosan to be used as oral immunoadjuvant for vaccination purposes. LBG-based nanoparticles were prepared by mild polyelectrolyte complexation between chitosan (CS) and a synthesized LBG sulfate derivative (LBGS). Morphological characterization suggested that nanoparticles present a solid and compact structure with spherical-like shape. Sizes around 180-200nm and a positive surface charge between +9mV and +14mV were obtained. CS/LBGS nanoparticles did not affect cell viability of Caco-2 cells after 3h and 24h of exposure when tested at concentrations up to 1.0mg/mL. Two model antigens (a particulate acellular extract HE of Salmonella enterica serovar Enteritidis, and ovalbumin as soluble antigen) were associated to CS/LBGS nanoparticles with efficiencies around 26% for ovalbumin and 32% for HE, which resulted in loading capacities up to 12%. The process did not affect the antigenicity of the associated antigens. BALB/c mice were orally immunized with ovalbumin-loaded nanoparticles (100μg), and results indicate an adjuvant effect of the CS/LBGS nanoparticles, eliciting a balanced Th1/Th2 immune response. Thus, CS/LBGS nanoparticles are promising as antigen mucosal delivery strategy, with particular interest for oral administration.