Differential glycosylation in the envelope glycoprotein enhances DC-SIGN-mediated cell entry of the West African Ebolavirus Makona.

During the 2013-2016 West African Ebola virus (EBOV) Makona epidemic, genomic surveillance identified an A82V substitution in the viral glycoprotein (GP) early in the outbreak. This mutation was associated with increased infectivity and faster viral spread, yet the host factors underlying this enhanced transmission remained unclear. Using a pseudotyped lentiviral system, we show that GP-A82V increases viral binding and infectivity in dendritic cells (DCs) and macrophages, key early and sustained target cells. This enhancement depends on expression of DC-SIGN, a lectin receptor on DCs. Filovirus-like particles carrying GP-V82 displayed accelerated infection kinetics in DC-SIGN-expressing cells. We identified two N-linked glycosylation sites, Ngly257 and Ngly563, that enhanced GP-V82 infectivity through simultaneous engagement of a DC-SIGN dimer. The A82V substitution increased soluble GP binding to DC-SIGN. Nuclear magnetic resonance analysis revealed enrichment of Lewis X (CD15), a known DC-SIGN ligand, in GP-V82. Blocking Lewis X with an anti-CD15 antibody reduced pseudovirus infection, confirming its functional role. These findings identify DC-SIGN as a key host factor contributing to EBOV Makona transmission and highlight viral envelope glycosylation as a mechanism facilitating viral adaptation and enhanced human-to-human spread.