title: Tetravalent neutralizing antibody response against four dengue serotypes by a single chimeric dengue envelope antigen.
abstract: We employed DNA shuffling and screening technologies to develop a single recombinant dengue envelope (E) antigen capable of inducing neutralizing antibodies against all four antigenically distinct dengue serotypes. By DNA shuffling of codon-optimized dengue 1-4 E genes, we created a panel of novel chimeric clones expressing C-terminal truncated E antigens that combined epitopes from all four dengue serotypes. DNA vaccines encoding these novel chimeras induced multivalent T cell and neutralizing antibody responses against all four dengue serotypes in mice. By contrast, a mixture of four unshuffled, parental DNA vaccines failed to produce tetravalent neutralizing antibodies in mice. The neutralizing antibody titers for some of these antigens could be further improved by extending the sequences to express full-length pre-membrane and envelope proteins. The chimeric antigens also protected mice against a lethal dengue-2 virus challenge. These data demonstrate that DNA shuffling and associated screening can lead to the selection of multi-epitope antigens against closely related dengue virus serotypes and suggest a broad utility for these technologies in optimizing vaccine antigens.
authors.affiliation: Maxygen Inc., Human Therapeutics, 515 Galveston Drive, Redwood City, CA 94063, USA. doris.apt@maxygen.com
authors.forename: Doris
authors.initials: D
authors.lastname: Apt
authors.forename: Kanakatte
authors.initials: K
authors.lastname: Raviprakash
authors.forename: Alice
authors.initials: A
authors.lastname: Brinkman
authors.forename: Andrey
authors.initials: A
authors.lastname: Semyonov
authors.forename: Shumin
authors.initials: S
authors.lastname: Yang
authors.forename: Craig
authors.initials: C
authors.lastname: Skinner
authors.forename: Lori
authors.initials: L
authors.lastname: Diehl
authors.forename: Richard
authors.initials: R
authors.lastname: Lyons
authors.forename: Kevin
authors.initials: K
authors.lastname: Porter
authors.forename: Juha
authors.initials: J
authors.lastname: Punnonen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D003715:Dengue / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D003716:Dengue Virus / Q000276:immunology*; D019020:Directed Molecular Evolution; D005434:Flow Cytometry; D015723:Gene Library; D007371:Interferon-gamma; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014759:Viral Envelope Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; D007371:Interferon-gamma