title: Mucosal vaccine targeting improves onset of mucosal and systemic immunity to botulinum neurotoxin A.
abstract: Absence of suitable mucosal adjuvants for humans prompted us to consider alternative vaccine designs for mucosal immunization. Because adenovirus is adept in binding to the respiratory epithelium, we tested the adenovirus 2 fiber protein (Ad2F) as a potential vaccine-targeting molecule to mediate vaccine uptake. The vaccine component (the host cell-binding domain to botulinum toxin (BoNT) serotype A) was genetically fused to Ad2F to enable epithelial binding. The binding domain for BoNT was selected because it lies within the immunodominant H chain as a beta-trefoil (Hcbetatre) structure; we hypothesize that induced neutralizing Abs should be protective. Mice were nasally immunized with the Hcbetatre or Hcbetatre-Ad2F, with or without cholera toxin (CT). Without CT, mice immunized with Hcbetatre produced weak secretory IgA (sIgA) and plasma IgG Ab response. Hcbetatre-Ad2F-immunized mice produced a sIgA response equivalent to mice coimmunized with CT. With CT, Hcbetatre-Ad2F-immunized mice showed a more rapid onset of sIgA and plasma IgG Ab responses that were supported by a mixed Th1/Th2 cells, as opposed to mostly Th2 cells by Hcbetatre-dosed mice. Mice immunized with adjuvanted Hcbetatre-Ad2F or Hcbetatre were protected against lethal BoNT serotype A challenge. Using a mouse neutralization assay, fecal Abs from Hcbetatre-Ad2F or Hcbetatre plus CT-dosed mice could confer protection. Parenteral immunization showed that the inclusion of Ad2F enhances anti-Hcbetatre Ab titers even in the absence of adjuvant. This study shows that the Hcbetatre structure can confer protective immunity and that use of Hcbetatre-Ad2F gives more rapid and sustained mucosal and plasma Ab responses.
authors.affiliation: Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717-3610, USA.
authors.forename: Massimo
authors.initials: M
authors.lastname: Maddaloni
authors.forename: Herman F
authors.initials: HF
authors.lastname: Staats
authors.forename: Dagmara
authors.initials: D
authors.lastname: Mierzejewska
authors.forename: Teri
authors.initials: T
authors.lastname: Hoyt
authors.forename: Amy
authors.initials: A
authors.lastname: Robinson
authors.forename: Gayle
authors.initials: G
authors.lastname: Callis
authors.forename: Shunji
authors.initials: S
authors.lastname: Kozaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kiyono
authors.forename: Jerry R
authors.initials: JR
authors.lastname: McGhee
authors.forename: Kohtaro
authors.initials: K
authors.lastname: Fujihashi
authors.forename: David W
authors.initials: DW
authors.lastname: Pascual
mesh_terms: D000276:Adjuvants, Immunologic; D000281:Administration, Intranasal; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects*; D000956:Antigens, Viral / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D001665:Binding Sites; D019274:Botulinum Toxins, Type A / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D002772:Cholera Toxin; D007143:Immunoglobulin Heavy Chains; D051379:Mice; D023421:Models, Animal; D009092:Mucous Membrane; D011993:Recombinant Fusion Proteins; D014612:Vaccines / Q000008:administration & dosage / Q000737:chemistry*
chemical_list: D000276:Adjuvants, Immunologic; D000956:Antigens, Viral; D036022:Capsid Proteins; D007143:Immunoglobulin Heavy Chains; D011993:Recombinant Fusion Proteins; D014612:Vaccines; C018315:hexon capsid protein, Adenovirus; D002772:Cholera Toxin; D019274:Botulinum Toxins, Type A