Cellobiose-coated Poly(lactide-Co-glycolide) Particles Loaded with Diphtheria Toxoid for Per Os Immunization.

Aim To evaluate the dose-dependent immunogenic properties of poly(lactide-co-glycolide) (PLGA) particles coated with cellobiose as antigen carriers for oral immunization.Methods Two types of PLGA-cellobiose particles (PLGA-cellobiose-1, similar to 0.8 mu m and PLGA-cellobiose-2, similar to 1.2 mu m) containing non-toxic recombinant subunit B (SbB) of diphtheria toxin fused with enhanced green fluorescent protein were characterized in vitro for their size, shape, antigen loading, and ability to induce phagocytosis. Different doses of antigen, immobilized on the particles (2.5 mu g, 25 mu g, 250 mu g, and 2500 mu g per 1 kg of body weight), were administered per os 3 times with intervals of 2 weeks to BALB/c female mice. The antigen-specific IgG and IgA antibodies were estimated in serum by ELISA.Results After the first immunization, increase in concentration of blood antitoxic antibodies was detected. Antigen dose 250 mu g/kg was the most immunogenic for IgG antibodies induction for both types of PLGA-cellobiose particles. Antigen doses 25 mu g/kg and 2.5 mu g/kg were the most immunogenic for IgA antibodies induction by PLGA-cellobiose 1 and 2 particles, respectively. The second and the third treatment had no significant effect on the immune response or even reduced it, which could be explained by immune tolerance induction by the antigens delivered per os.Conclusion Our results suggest that the correct dose of PLGA-cellobiose particles loaded with antigen could significantly increase the humoral immune response against the introduced antigen already after the first immunization. Thus, PLGA particles can be considered as a potent component of oral vaccines.