Enhanced Enterovirus D68 Replication in Neuroblastoma Cells Is Associated with a Cell Culture-Adaptive Amino Acid Substitution in VP1.

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dc.contributor.authorSooksawasdi Na Ayudhya, Syriam
dc.contributor.authorMeijer, Adam
dc.contributor.authorBauer, Lisa
dc.contributor.authorOude Munnink, Bas
dc.contributor.authorEmbregts, Carmen
dc.contributor.authorLeijten, Lonneke
dc.contributor.authorSiegers, Jurre Y
dc.contributor.authorLaksono, Brigitta M
dc.contributor.authorvan Kuppeveld, Frank
dc.contributor.authorKuiken, Thijs
dc.contributor.authorGeurts-van Kessel, Corine
dc.contributor.authorvan Riel, Debby
dc.contributor.authorLakdawala, Seema
dc.date.accessioned2020-11-11T13:09:29Z
dc.date.available2020-11-11T13:09:29Z
dc.date.issued2020-11-04
dc.description.abstractSince its emergence in the United States in 2014, enterovirus D68 (EV-D68) has been and is associated with severe respiratory diseases and acute flaccid myelitis. Even though EV-D68 has been shown to replicate in different neuronal cells in vitro, it is currently poorly understood which viral factors contribute to the ability to replicate efficiently in cells of the central nervous system and whether this feature is a clade-specific feature. Here, we determined the replication kinetics of clinical EV-D68 isolates from (sub)clades A, B1, B2, B3, and D1 in human neuroblastoma cells (SK-N-SH). Subsequently, we compared sequences to identify viral factors associated with increased viral replication. All clinical isolates replicated in SK-N-SH cells, although there was a large difference in efficiency. Efficient replication of clinical isolates was associated with an amino acid substitution at position 271 of VP1 (E271K), which was acquired during virus propagation in vitro Recognition of heparan sulfate in addition to sialic acids was associated with increased attachment, infection, and replication. Removal of heparan sulfate resulted in a decrease in attachment, internalization, and replication of viruses with E271K. Taken together, our study suggests that the replication kinetics of EV-D68 isolates in SK-N-SH cells is not a clade-specific feature. However, recognition of heparan sulfate as an additional receptor had a large effect on phenotypic characteristics in vitro. These observations emphasize the need to compare sequences from virus stocks with clinical isolates in order to retrieve phenotypic characteristics from original virus isolates.IMPORTANCE Enterovirus D68 (EV-D68) causes mild to severe respiratory disease and is associated with acute flaccid myelitis since 2014. Currently, the understanding of the ability of EV-D68 to replicate in the central nervous system (CNS), and whether it is associated with a specific clade of EV-D68 viruses or specific viral factors, is lacking. Comparing different EV-D68 clades did not reveal clade-specific phenotypic characteristics. However, we did show that viruses which acquired a cell culture-adapted amino acid substitution in VP1 (E271K) recognized heparan sulfate as an additional receptor. Recognition of heparan sulfate resulted in an increase in attachment, infection, and replication in neuroblastoma cells compared with viruses without this specific amino acid substitution. The ability of EV-D68 viruses to acquire cell culture-adaptive substitutions which have a large effect in experimental settings emphasizes the need to sequence virus stocks.en_US
dc.identifier.doi10.1128/mSphere.00941-20
dc.identifier.issn2379-5042
dc.identifier.journalmSphere 2020; 5(6):e00941en_US
dc.identifier.pmid33148825
dc.identifier.urihttp://hdl.handle.net/10029/624472
dc.language.isoenen_US
dc.source.journaltitlemSphere
dc.subjectVP1en_US
dc.subjectcell culture adaptationen_US
dc.subjectenterovirus D68en_US
dc.subjectheparan sulfateen_US
dc.subjectin vitroen_US
dc.subjectneuroblastoma cellsen_US
dc.subjectneurotropismen_US
dc.subjectpathogenesisen_US
dc.subjectreplicationen_US
dc.titleEnhanced Enterovirus D68 Replication in Neuroblastoma Cells Is Associated with a Cell Culture-Adaptive Amino Acid Substitution in VP1.en_US
dc.typeArticleen_US
dspace.entity.typePublication
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