Abstract
Ebolaviruses continue to pose a significant outbreak threat, and while Ebola virus (EBOV)-specific vaccines and antivirals have been licensed, efforts to develop candidates offering broad species cross-protection are continuing. The use of pseudotyped virus in place of live virus is recognised as an alternative, safer, high-throughput platform to evaluate anti-ebolavirus antibodies towards their development, yet it requires optimisation. Here, we have shown that the target cell line impacts neutralisation assay results and cannot be selected purely based on permissiveness. In expanding the platform to incorporate each of the ebolavirus species envelope glycoprotein, allowing a comprehensive assessment of cross-neutralisation, we found that the recently discovered Bombali virus has a point mutation in the receptor-binding domain which prevents entry into a hamster cell line and, importantly, shows that this virus can be cross-neutralised by EBOV antibodies and convalescent plasma.
Highlights
Introduction28,000 cases and and health emergency of international concern (PHEIC) and caused over 28,000 cases
The 2013–2016 outbreak of Ebola virusvirus in WestAfrica, which was declared a public outbreak of Ebola in WestAfrica, which was declared a public emergency of international concern (PHEIC) and caused over28,000 cases and and health emergency of international concern (PHEIC) and caused over 28,000 casesPublished: August with24 regard to jurisdictional claims 11,000 in deaths [1,2],[1,2], re-focused thethe world’s attention ononthe theEbolavirusEbo11,000 deaths re-focused world’s attention thepublic publichealth health threat threat the published maps and institutional affillavirus genus represents.This included the establishment and inclusion ofEbola virus genus represents
Culture medium was removed and the target cells were incubated for 5 mins with a 1:1 mix of phenol-free DMEM (Gibco) and Bright-Glo reagent (Promega), with luciferase activity detected as relative light units (RLU) using a Glomax
Summary
28,000 cases and and health emergency of international concern (PHEIC) and caused over 28,000 cases. In 2018, as part of the PREDICT project to detect known and unknown viruses within wildlife reservoirs [11,12,13], the genome sequence of a further ebolavirus, Bombali virus (BOMV), was isolated from free-tailed bats in Sierra Leone [14]. In light of the disease potential (i.e., documented human cases or ability to infect human cells in vitro) posed by all 6 Ebolavirus species, there are efforts to develop cross-protective candidates [21]. To evaluate their effectiveness and further study serological responses against existing and newly discovered ebolaviruses, species-specific neutralisation assays are required. We have optimised a lentiviral-vector based (LVV) neutralisation assay for EBOV antibodies and applied it in the evaluation of cross-neutralising antibodies against the other ebolavirus species, including BOMV
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