Elsevier

Virology

Volume 499, December 2016, Pages 350-360
Virology

Mutations in VP1 and 3A proteins improve binding and replication of rhinovirus C15 in HeLa-E8 cells

https://doi.org/10.1016/j.virol.2016.09.025Get rights and content
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Highlights

  • We adapted RV-C15 clinical isolate to efficient growth in HeLa-E8 cells.

  • Serial passaging resulted in stronger cytopathic effects and increased virus binding to cells and progeny yields.

  • We identified two key mutations which increased virus binding (VP1 T125K) and replication (3A E41K), respectively

  • The findings enabled large-scale cost-effective C15 production by infection and the testing of RV-C infectivity by plaque assay.

Abstract

Viruses in the rhinovirus C species (RV-C) can cause severe respiratory illnesses in children including pneumonia and asthma exacerbations. A transduced cell line (HeLa-E8) stably expressing the CDHR3-Y529 receptor variant, supports propagation of RV-C after infection. C15 clinical or recombinant isolates replicate in HeLa-E8, however progeny yields are lower than those of related strains of RV-A and RV-B. Serial passaging of C15 in HeLa-E8 resulted in stronger cytopathic effects and increased (≥10-fold) virus binding to cells and progeny yields. The adaptation was acquired by two mutations which increased binding (VP1 T125K) and replication (3A E41K), respectively. A similar 3A mutation engineered into C2 and C41 cDNAs also improved viral replication (2–8 fold) in HeLa but the heparan sulfate mediated cell-binding enhancement by the VP1 change was C15-specific. The findings now enable large-scale cost-effective C15 production by infection and the testing of RV-C infectivity by plaque assay.

Keywords

Rhinovirus C
Pathogenesis
Adaptation
Receptor specificity
Heparan sulfate

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