title: Assessment of DNA vaccine potential for gilthead sea bream (Sparus aurata) by intramuscular injection of a reporter gene.
abstract: Naked circular plasmid DNA containing the cytomegalovirus (CMV)-promoter-driven lacZ reporter gene (pCMV-LacZ) was injected in the epaxial muscle of gilthead sea bream (Sparus aurata). A mosaic pattern of expression of beta-galactosidase (beta-gal) in the myofibres at the site of injection was visualised by in situ histochemical staining using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside. As measured by o-nitrophenyl-beta-D-galactopyranoside assay, beta-gal enzymatic activity was found to steadily increase for at least 50 days post injection (p.i.) in pCMV-LacZ-injected muscle. In parallel, foreign DNA was detected by polymerase chain reaction in injected muscles (but not in other tissues) up to 60 days p.i., persisting most probably in an extrachromosomal, non-replicative, circular form. Neither beta-gal activity nor pCMV-LacZ-related amplification products were found 90 days p.i. Antibodies against beta-gal were demonstrated in pCMV-LacZ-injected fish sampled 45 days p.i. The results suggest that intramuscular delivery of foreign genes represents a realistic approach for DNA vaccine technology for the prevention of infectious diseases in gilthead sea bream.
authors.affiliation: Department of Biological and Environmental Sciences and Technologies, University of Lecce, via Provinciale Lecce-Monteroni, I-73100, Lecce, Italy. physiol@ultra5.unile.it
authors.forename: Tiziano
authors.initials: T
authors.lastname: Verri
authors.forename: Laura
authors.initials: L
authors.lastname: Ingrosso
authors.forename: Rita
authors.initials: R
authors.lastname: Chiloiro
authors.forename: Antonio
authors.initials: A
authors.lastname: Danieli
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Zonno
authors.forename: Pietro
authors.initials: P
authors.lastname: Alifano
authors.forename: Nicla
authors.initials: N
authors.lastname: Romano
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Scapigliati
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Vilella
authors.forename: Carlo
authors.initials: C
authors.lastname: Storelli
mesh_terms: D000818:Animals; D017756:Aquaculture / Q000379:methods; D005393:Fish Diseases / Q000517:prevention & control*; D015870:Gene Expression; D017930:Genes, Reporter / Q000276:immunology*; D006652:Histological Techniques; D007273:Injections, Intramuscular; D007763:Lac Operon / Q000276:immunology; D016133:Polymerase Chain Reaction; D021541:Sea Bream / Q000276:immunology*; D014611:Vaccination / Q000662:veterinary*; D019444:Vaccines, DNA / Q000276:immunology*; D001616:beta-Galactosidase / Q000276:immunology / Q000378:metabolism
chemical_list: D019444:Vaccines, DNA; D001616:beta-Galactosidase