title: Construction and characterization of E1- and E3-deleted adenovirus vectors expressing two antigens from two separate expression cassettes.
abstract: Here we describe a series of replication-defective adenovirus vectors designed to express transgene products from two expression cassettes placed into the deleted E1 and E3 domains. Vectors that contained an E1 cassette with a cytomegalovirus promoter in the forward orientation and an E3 cassette with the chicken β-actin promoter in the reverse orientation grew to acceptable yields and expressed both transgenes. Additionally, they elicited immune responses to both transgene products. Levels of expression and the vectors' immunogenicity were influenced by the presence of regulatory elements shared between the two expression cassettes. Specifically, vectors that carried the same intron and enhancer in both expression cassettes could be rescued and expanded, but they were poorly immunogenic. Deletion of the enhancer or both the enhancer and the intron from the E3 cassette increased T- and B-cell responses to both transgene products.
authors.affiliation: 1 Cellular and Molecular Biology, University of Pennsylvania , Philadelphia, PA 19104.
authors.forename: Juliana C
authors.initials: JC
authors.lastname: Small
authors.forename: Raj K
authors.initials: RK
authors.lastname: Kurupati
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Zhou
authors.forename: Ang
authors.initials: A
authors.lastname: Bian
authors.forename: Emily
authors.initials: E
authors.lastname: Chi
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Zhiquan
authors.initials: Z
authors.lastname: Xiang
authors.forename: Hildegund C J
authors.initials: HC
authors.lastname: Ertl
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology; D017498:Adenovirus E1 Proteins / Q000235:genetics*; D017503:Adenovirus E3 Proteins / Q000235:genetics*; D000818:Animals; D000941:Antigens / Q000235:genetics* / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D005260:Female; D017353:Gene Deletion; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics* / Q000276:immunology; D016679:Genome, Viral; D042822:Genomic Instability; D006801:Humans; D051379:Mice; D019076:Transgenes / Q000235:genetics* / Q000276:immunology
chemical_list: D017498:Adenovirus E1 Proteins; D017503:Adenovirus E3 Proteins; D000941:Antigens