title: Quantitative adenovirus neutralization assays based on the secreted alkaline phosphatase reporter gene: application in epidemiologic studies and in the design of adenovector vaccines.
abstract: Replication-defective recombinant adenoviruses (rAd) are used as vectors for vaccines as well as for gene therapy. To determine type-specific antibodies to adenovirus (Ad) serotypes 2, 5, 24, 34, and 35, we developed quantitative neutralization assays using recombinant adenoviruses with the secreted alkaline phosphatase (SEAP) reporter gene. Among the standardized parameters, the concentration of infectious and noninfectious adenoviral particles used in the assay is critical for a reliable comparison of data from different studies. The usefulness of this assay was demonstrated in a pilot epidemiologic study of 40 healthy individuals. In this study, the highest prevalence of antiadenovirus antibodies was found for the Ad2 serotype (82.5%), followed by Ad5 (35%). The prevalence of antiadenovirus antibodies for the serotypes 24, 34, and 35 was low (7.5%, 2.5%, and 0%, respectively). In addition, epidemiologic parameters such as gender and age were statistically evaluated. A positive association was found between age and the presence of anti-Ad5 antibodies. The assay was also useful for evaluating the presence of antiadenovirus antibodies in the design of vaccines using a rhesus monkey model. In this animal model, it was possible to determine differential dose and time responses, and the specificity for the detection of neutralizing antibodies was assessed. The evaluation of serotype-specific neutralizing antibodies can be of both clinical and epidemiologic importance as a means of selecting the appropriate serotype adenovector(s).
authors.affiliation: Department of Virus & Cell Biology, Merck Research Laboratories, Merck & Co., Inc., West Point, PA 19486, USA.
authors.forename: Miguel
authors.initials: M
authors.lastname: Aste-Amézaga
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Bett
authors.forename: Fubao
authors.initials: F
authors.lastname: Wang
authors.forename: Danilo R
authors.initials: DR
authors.lastname: Casimiro
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Antonello
authors.forename: Deepa K
authors.initials: DK
authors.lastname: Patel
authors.forename: Elayne C
authors.initials: EC
authors.lastname: Dell
authors.forename: Laura L
authors.initials: LL
authors.lastname: Franlin
authors.forename: Nancy M
authors.initials: NM
authors.lastname: Dougherty
authors.forename: Philip S
authors.initials: PS
authors.lastname: Bennett
authors.forename: Helen C
authors.initials: HC
authors.lastname: Perry
authors.forename: Mary-Ellen
authors.initials: ME
authors.lastname: Davies
authors.forename: John W
authors.initials: JW
authors.lastname: Shiver
authors.forename: Paul M
authors.initials: PM
authors.lastname: Keller
authors.forename: Mark D
authors.initials: MD
authors.lastname: Yeager
mesh_terms: D000256:Adenoviridae / Q000145:classification / Q000235:genetics* / Q000276:immunology*; D000328:Adult; D000368:Aged; D000469:Alkaline Phosphatase / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000276:immunology; D002460:Cell Line; D016021:Epidemiologic Studies; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000276:immunology*; D006801:Humans; D008253:Macaca mulatta; D008297:Male; D008875:Middle Aged; D009500:Neutralization Tests / Q000379:methods*; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D019444:Vaccines, DNA; D000469:Alkaline Phosphatase