Tetravalent neutralizing antibody response against four dengue serotypes by a single chimeric dengue envelope antigen

• Published in: Vaccine 2006-01, Vol.24 (3), p.335-344
• Main Authors: Apt, Doris, Raviprakash, Kanakatte, Brinkman, Alice, Semyonov, Andrey, Yang, Shumin, Skinner, Craig, Diehl, Lori, Lyons, Richard, Porter, Kevin, Punnonen, Juha
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Viral - biosynthesis; Antibodies, Viral - immunology; Antigen; Applied microbiology; Arboviroses; Biological and medical sciences; Blotting, Western; Cell Line; Cloning; Cross-neutralizing antibodies; Dengue - immunology; Dengue - prevention & control; Dengue - virology; Dengue fever; Dengue fevers; Dengue virus; Dengue Virus - immunology; Dengue virus type 2; Deoxyribonucleic acid; Directed Molecular Evolution; DNA; DNA shuffling; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Gene Library; Human viral diseases; Infections; Infectious diseases; Interferon-gamma; Laboratory animals; Medical sciences; Mice; Mice, Inbred BALB C; Microbiology; Miscellaneous; Molecular weight; Multivalent antigen; Neutralization Tests; Plasmids - genetics; Proteins; Recombinant Fusion Proteins - immunology; T-Lymphocytes - immunology; Tetravalent; Transfection; Tropical diseases; Tropical viral diseases; Vaccines; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects); Vaccines, DNA - immunology; Vaccines, Synthetic - immunology; Vector-borne diseases; Viral diseases; Viral Envelope Proteins - immunology; Viral Vaccines - immunology; Virology; Viruses
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2005.07.100

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.