title: Stimulation of IgY responses in gene gun immunized laying hens by combined administration of vector DNA coding for the target antigen Botulinum toxin A1 and for avian cytokine adjuvants.
abstract: DNA immunization is a convenient and effective way of inducing a specific antibody response. In mammals, co-administration of vectors encoding immunostimulatory cytokines can enhance the humoral response resulting in elevated antibody titers. We therefore set out to investigate the effect using avian interleukin 1β (IL-1β) and avian interleukin 6 (IL-6) as genetic adjuvants when immunizing laying hens. A BoNT A1 holotoxoid DNA immunogen carrying two inactivating mutations was evaluated for its ability to induce a specific and sustained IgY antibody response. Both the holotoxoid and the cytokine sequences were codon-optimized. In vitro, the proteins were efficiently expressed in transfected HEK 293T cells and the cytokines were secreted into the culture supernatants. Whereas eggs from hens immunized via gene gun using a prime boost strategy showed no differences in their total IgY content, the specific αBoNT A1 response was slightly elevated up to 1.4× by the IL-1β adjuvant vector and increased by 3.8× by the IL-6 vector. Finally, although hens receiving the IL-1β adjuvant had laying capacities above the average, hens receiving the IL-6 adjuvant experienced laying problems.
authors.affiliation: FU-Berlin, Fachbereich Biologie, Chemie, Pharmazie, Berlin, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Niederstadt
authors.forename: Oliver
authors.initials: O
authors.lastname: Hohn
authors.forename: Brigitte G
authors.initials: BG
authors.lastname: Dorner
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Schade
authors.forename: Norbert
authors.initials: N
authors.lastname: Bannert
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D019470:Biolistics / Q000379:methods*; D019274:Botulinum Toxins, Type A / Q000235:genetics / Q000276:immunology*; D002645:Chickens / Q000235:genetics / Q000276:immunology*; D016207:Cytokines / Q000276:immunology*; D004247:DNA / Q000235:genetics* / Q000276:immunology; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D007136:Immunoglobulins / Q000276:immunology*; D053583:Interleukin-1beta / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; C016512:IgY; D007136:Immunoglobulins; D053583:Interleukin-1beta; D015850:Interleukin-6; D004247:DNA; D019274:Botulinum Toxins, Type A