caa a22 4500 445311568 CHVBK 20180317142631.0 cr unu---uuuuu 170323e20110101xx s 000 0 eng 10.1007/s11095-010-0093-y doi (NATIONALLICENCE)springer-10.1007/s11095-010-0093-y Transcutaneous Immunization Studies in Mice Using Diphtheria Toxoid-Loaded Vesicle Formulations and a Microneedle Array [Elektronische Daten] [Zhi Ding, Suzanne Bal, Stefan Romeijn, Gideon Kersten, Wim Jiskoot, Joke Bouwstra] ABSTRACT: Purpose: To determine the immunogenicity of diphtheria toxoid (DT) formulated in two types of vesicles following transcutaneous immunization (TCI) of mice onto microneedle array-treated skin. Methods: DT-containing cationic liposomes or anionic surfactant-based vesicles were prepared by extrusion and sonication. The physicochemical properties were characterized in terms of size, ζ-potential, vesicle elasticity and antigen association. TCI was performed by applying formulations onto intact or microneedle array-pretreated mice skin, using cholera toxin as an adjuvant. Subcutaneous and intradermal immunizations were as control. Immune responses were evaluated by IgG and neutralizing antibody titers, and the immune-stimulatory properties were assessed using cultured dendritic cells. Results: Stable DT-containing cationic liposomes (∼150nm) and anionic vesicles (∼100nm) were obtained. Incorporation of Span 80 increased liposome elasticity. About 90% and 77% DT was associated with liposomes and vesicles, respectively. TCI of all formulations resulted in substantial antibody titers only if microneedle pretreatment was applied. Co-administration of cholera toxin further augmented the immune responses of TCI. However, vesicle formulations didn't enhance the immunogenicity on either intact or microneedle-treated skin and showed low stimulatory activity on dendritic cells. Conclusions: Microneedle pretreatment and cholera toxin, but not antigen association to vesicles, enhances the immunogenicity of topically applied DT. The Author(s), 2010 cholera toxin nationallicence diphtheria toxoid nationallicence microneedle array nationallicence transcutaneous immunization nationallicence vesicles nationallicence Alum : Adju-Phos®, colloidal aluminum phosphate nationallicence APCs : antigen-presenting cells nationallicence CB(S) : citrate buffer(ed saline) nationallicence CT : cholera toxin nationallicence DC : dendritic cell nationallicence DOTAP : 1,2-dioleoyl-3-trimethylammonium-propane chloride salt nationallicence DT : diphtheria toxoid nationallicence GM-CSF : granulocyte-macrophage colony-stimulating factor nationallicence HRP-GAM : horseradish peroxidase conjugated goat anti-mouse nationallicence i.d. : intradermal nationallicence IL-4 : interleukin-4 nationallicence L-595 : sucrose-laurate ester nationallicence LPS : lipopolysaccharide nationallicence PB(S) : phosphate buffer(ed saline) nationallicence PDI : polydispersity index nationallicence PEG-8-L : octaoxyethylene laurate ester nationallicence s.c. : subcutaneous nationallicence SEC : size exclusion chromatography nationallicence SPC : soybean phosphatidyl choline nationallicence TCI : transcutaneous immunization nationallicence TMB : chromogen 3, 3′, 5, 5′-tetramethylbenzidine nationallicence TR-70 : sodium bistridecyl sulfo succinate nationallicence Ding Zhi Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut Bal Suzanne Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut Romeijn Stefan Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut Kersten Gideon Department of Research and Development, Netherlands Vaccine Institute, 3720 BA, Bilthoven, The Netherlands aut Jiskoot Wim Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut Bouwstra Joke Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/1(2011-01-01), 145-158 0724-8741 28:1<145 2011 28 11095 https://doi.org/10.1007/s11095-010-0093-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-010-0093-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ding Zhi Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Bal Suzanne Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Romeijn Stefan Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Kersten Gideon Department of Research and Development, Netherlands Vaccine Institute, 3720 BA, Bilthoven, The Netherlands aut NATIONALLICENCE 700 1- Jiskoot Wim Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Bouwstra Joke Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/1(2011-01-01), 145-158 0724-8741 28:1<145 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer