title: Generation of a novel replication-incompetent adenoviral vector derived from human adenovirus type 49: manufacture on PER.C6 cells, tropism and immunogenicity.
abstract: Recombinant adenoviral vectors based on type 5 (rAd5) show great promise as a vaccine carrier. However, neutralizing activity against Ad5 is prevalent and high-titred among human populations, and significantly dampens Ad5-based vaccine modalities. The generation of alternative adenoviral vectors with low seroprevalence thus receives much research attention. Here, it is shown that a member from human adenovirus subgroup D, i.e. Ad49, does not cross-react with Ad5 neutralizing activity, making it a candidate serotype for vector development. Therefore, a plasmid system that allows formation of replication-incompetent adenovirus serotype 49 vaccine vectors (rAd49) was constructed and it was demonstrated that rAd49 can be successfully propagated to high titres on existing Ad5.E1-complementing cell lines such as PER.C6. Using an rAd49 vector carrying the luciferase marker gene, detailed seroprevalence studies were performed, demonstrating that rAd49 has low seroprevalence and neutralizing antibody titres worldwide. Also, we have initiated rAd49 vector receptor usage suggesting that rAd49 utilizes hCD46 as a cellular receptor. Finally, the immunogenicity of the rAd49 vector was assessed and it was shown that an rAd49.SIVGag vaccine induces strong anti-SIVGag CD8+ T-lymphocytes in naïve mice, albeit less than an rAd5.SIVGag vaccine. However, in mice with high anti-Ad5 immunity the rAd5.SIVGag vaccine was severely blunted, whereas the anti-SIVGag response was not significantly suppressed using the rAd49.SIVGag vaccine. These data demonstrate the potential of a replication deficient human group D adenoviral vector for vaccination purposes.
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Angelique A C
authors.initials: AAC
authors.lastname: Lemckert
authors.affiliation: Gaubius Laboratory, TNO, 2301 CA Leiden, The Netherlands.
authors.forename: Jos
authors.initials: J
authors.lastname: Grimbergen
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Shirley
authors.initials: S
authors.lastname: Smits
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Eric
authors.initials: E
authors.lastname: Hartkoorn
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Lennart
authors.initials: L
authors.lastname: Holterman
authors.affiliation: Department of Human Retrovirology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Ben
authors.initials: B
authors.lastname: Berkhout
authors.affiliation: Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, MA 02215, USA.
authors.forename: Dan H
authors.initials: DH
authors.lastname: Barouch
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Ronald
authors.initials: R
authors.lastname: Vogels
authors.affiliation: Department of Surgery, LUMC, Leiden, The Netherlands.|Gaubius Laboratory, TNO, 2301 CA Leiden, The Netherlands.
authors.forename: Paul
authors.initials: P
authors.lastname: Quax
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Jaap
authors.initials: J
authors.lastname: Goudsmit
authors.affiliation: Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
authors.forename: Menzo J E
authors.initials: MJE
authors.lastname: Havenga
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics / Q000254:growth & development* / Q000276:immunology* / Q000502:physiology; D000818:Animals; D000914:Antibodies, Viral; D002460:Cell Line; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006801:Humans; D051916:Membrane Cofactor Protein; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009928:Organ Specificity; D014614:Vaccines, Synthetic; D014765:Viral Vaccines / Q000276:immunology; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D051916:Membrane Cofactor Protein; D014614:Vaccines, Synthetic; D014765:Viral Vaccines