An Engineered N-Glycosylated Dengue Envelope Protein Domain III Facilitates Epitope-Directed Selection of Potently Neutralizing and Minimally Enhancing Antibodies.

Autor:	Nilchan N; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Kraivong R; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Luangaram P; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Phungsom A; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Tantiwatcharakunthon M; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Traewachiwiphak S; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Prommool T; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand., Punyadee N; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand.; Division of Dengue Hemorrhagic Fever Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand., Avirutnan P; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand.; Division of Dengue Hemorrhagic Fever Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand., Duangchinda T; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Medical Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand., Malasit P; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Siriraj Center of Research Excellence in Dengue and Emerging Pathogens Mahidol University, Bangkok 10700, Thailand.; Division of Dengue Hemorrhagic Fever Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand., Puttikhunt C; Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.; Medical Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.
Jazyk:	angličtina
Zdroj:	ACS infectious diseases [ACS Infect Dis] 2024 Aug 09; Vol. 10 (8), pp. 2690-2704. Date of Electronic Publication: 2024 Jun 29.
DOI:	10.1021/acsinfecdis.4c00058
Abstrakt:	The envelope protein of dengue virus (DENV) is a primary target of the humoral immune response. The domain III of the DENV envelope protein (EDIII) is known to be the target of multiple potently neutralizing antibodies. One such antibody is 3H5, a mouse antibody that binds strongly to EDIII and potently neutralizes DENV serotype 2 (DENV-2) with unusually minimal antibody-dependent enhancement (ADE). To selectively display the binding epitope of 3H5, we strategically modified DENV-2 EDIII by shielding other known epitopes with engineered N-glycosylation sites. The modifications resulted in a glycosylated EDIII antigen termed "EDIII mutant N". This antigen was successfully used to sift through a dengue-immune scFv-phage library to select for scFv antibodies that bind to or closely surround the 3H5 epitope. The selected scFv antibodies were expressed as full-length human antibodies and showed potent neutralization activity to DENV-2 with low or negligible ADE resembling 3H5. These findings not only demonstrate the capability of the N-glycosylated EDIII mutant N as a tool to drive an epitope-directed antibody selection campaign but also highlight its potential as a dengue immunogen. This glycosylated antigen shows promise in focusing the antibody response toward a potently neutralizing epitope while reducing the risk of antibody-dependent enhancement.
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu