Abstract
Dengue and Zika are two of the most important human viral pathogens worldwide. In both cases, the envelope glycoprotein E is the main target of the antibody response. Recently, new complex quaternary epitopes were identified which are the consequence of the arrangement of the antiparallel E dimers on the viral surface. Such epitopes can be exploited to develop more efficient cross-neutralizing vaccines. Here we describe a successful covalent stabilization of E dimers from Dengue and Zika viruses in mammalian cells. Folding and dimerization of secretory E was found to be strongly dependent on temperature but independent of PrM co-expression. In addition, we found that, due to the close relationship between flaviviruses, Dengue and Zika viruses E proteins can form heterodimers and assemble into mosaic viral particles. Finally, we present new virus-free analytical platforms to study and screen antibody responses against Dengue and Zika, which allow for differentiation of epitopes restricted to specific domains, dimers and higher order arrangements of E.
Highlights
The Flaviviridae family includes some of the most important arthropod-borne human pathogens such as Dengue virus (DENV), Zika virus (ZIKV), West Nile virus (WNV) and Yellow fever virus (YFV)[1]
Given that some of the most potent cross-neutralizing antibodies against DENV and ZIKV recognize complex quaternary epitopes that only exist within the context of the E dimer[28], we attempted to stabilize the dimeric conformation of recombinant E from DENV and ZIKV expressed in mammalian cells
We have previously reported a detailed analysis of the secretory profile of recombinant E derived from the different DENV serotypes in mammalian cells and shown that DENV2 E ectodomain (2sE) undergoes different folding depending on temperature[36]
Summary
The Flaviviridae family includes some of the most important arthropod-borne human pathogens such as Dengue virus (DENV), Zika virus (ZIKV), West Nile virus (WNV) and Yellow fever virus (YFV)[1]. On the surface of the mature viral particle, E folds into an elongated rod-like structure forming 90 antiparallel homodimers, organized in 30 rafts, each composed of 3 parallel E dimers distributed in a herringbone-like configuration[8]. The 4 loosely packed peptide strands that connect DI and DII form a functional domain named Hinge region, which provides the flexibility needed for E trimerization[14]. In this pH-induced conformational reorganization of the viral surface, DII drives E reorientation from a horizontal antiparallel dimer into a vertical parallel trimer exposing the FL outwards to initiate the fusion process[15]. Upon release from the cell, the neutral pH of the extracellular environment stabilizes E dimers and promotes dissociation of Pr18
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
