title: Chitosan microparticles for mucosal vaccination against diphtheria: oral and nasal efficacy studies in mice.
abstract: In this study, the ability of chitosan microparticles to enhance both the systemic and local immune responses against diphtheria toxoid (DT) after oral and nasal administration in mice was investigated.Firstly, DT was associated to chitosan microparticles to determine antigen loading and release. Then DT loaded chitosan microparticles, DT in phosphate buffered saline (PBS) and empty chitosan microparticles (as controls) were fed intragastrically and administered nasally to mice. Mice were also subcutaneously immunised with DT associated with alum. All mice were vaccinated in week 1 and boosted in week 3. Sera were analysed for anti-DT IgG and nasal washings and faeces for anti-DT IgA titres using an enzyme linked immunosorbent assay. Loading capacities of about 25% and loading efficacies of about 100% were obtained after loading the chitosan microparticles with DT. No DT was released at 37 degrees C in PBS. Compared to intragastrical feeding with DT in PBS, a strong enhancement of the systemic and local immune responses against DT were found in mice fed with DT loaded chitosan microparticles. In addition, a dose-dependent immune reaction was observed for mice vaccinated with different doses of DT associated to chitosan microparticles. Significant systemic humoral immune responses were also found after nasal vaccination with DT associated to chitosan microparticles.DT associated to chitosan microparticles results in protective systemic and local immune response against DT after oral vaccination, and in significant enhancement of IgG production after nasal administration. Hence, these in vivo experiments demonstrate that chitosan microparticles are very promising mucosal vaccine delivery systems.
authors.affiliation: Leiden/Amsterdam Center for Drug Research, Division of Pharmaceutical Technology, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
authors.forename: Inez M
authors.initials: IM
authors.lastname: van der Lubben
authors.forename: Gideon
authors.initials: G
authors.lastname: Kersten
authors.forename: Marjan M
authors.initials: MM
authors.lastname: Fretz
authors.forename: Coen
authors.initials: C
authors.lastname: Beuvery
authors.forename: J
authors.initials: J
authors.lastname: Coos Verhoef
authors.forename: Hans E
authors.initials: HE
authors.lastname: Junginger
mesh_terms: D000281:Administration, Intranasal; D000284:Administration, Oral; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D002686:Chitin / Q000008:administration & dosage* / Q000031:analogs & derivatives*; D048271:Chitosan; D004168:Diphtheria Toxoid / Q000008:administration & dosage* / Q000276:immunology; D005260:Female; D018928:Immunity, Mucosal; D007070:Immunoglobulin A / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D014611:Vaccination
chemical_list: D000907:Antibodies, Bacterial; D004168:Diphtheria Toxoid; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D002686:Chitin; D048271:Chitosan