title: Conjugation of ovalbumin to trimethyl chitosan improves immunogenicity of the antigen.
abstract: Subunit vaccines are generally safer, but often less effective than live attenuated vaccines as they lack the necessary co-stimulatory factors. The formulation of an adjuvant like N-trimethyl chitosan (TMC) with an antigen can overcome its poor immunogenicity. Recent data suggest the importance of incorporating the antigen and the adjuvant into one entity for maximum immunostimulatory effect, e.g. by using (nano)particles. In the present paper we introduce the conjugation of an antigen, ovalbumin (OVA), to TMC as an alternative to nanoparticles for subunit vaccination. OVA was covalently linked to TMC using thiol chemistry (SPDP method). The uptake of the resulting TMC-OVA conjugate by dendritic cells (DC) and its effect on DC maturation was assessed in vitro and its immunogenicity was investigated in mice. We found that with the SPDP method a reducible covalent bond between TMC and OVA could be introduced, without disrupting the protein's antigenicity and structure. Uptake of TMC-OVA conjugate by dendritic cells was similar to the uptake of TMC/OVA nanoparticles, over 5-fold increased compared to a solution of OVA and TMC. Mice immunized with TMC-OVA conjugate produced 1000-fold higher OVA specific IgG titers than mice immunized with either OVA or a physical mixture of TMC and OVA. Moreover, these antibody titers were slightly elevated compared to the titers obtained with TMC/OVA nanoparticles. Conjugation of the antigen to an adjuvant is therefore a viable strategy to increase the immunogenicity of subunit vaccines and may provide an alternative to the use of particles.
authors.affiliation: Division of Drug Delivery Technology, Leiden/Amsterdam Center for Drug Research, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Bram
authors.initials: B
authors.lastname: Slütter
authors.forename: Peter Christiaan
authors.initials: PC
authors.lastname: Soema
authors.forename: Zhi
authors.initials: Z
authors.lastname: Ding
authors.forename: Rolf
authors.initials: R
authors.lastname: Verheul
authors.forename: Wim
authors.initials: W
authors.lastname: Hennink
authors.forename: Wim
authors.initials: W
authors.lastname: Jiskoot
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry; D000818:Animals; D000941:Antigens / Q000737:chemistry* / Q000276:immunology*; D048271:Chitosan / Q000737:chemistry* / Q000276:immunology*; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry; D010047:Ovalbumin / Q000737:chemistry* / Q000276:immunology*; D014611:Vaccination
chemical_list: D000276:Adjuvants, Immunologic; D000941:Antigens; D007074:Immunoglobulin G; C118071:N-trimethyl chitosan chloride; D010047:Ovalbumin; D048271:Chitosan