Mutations in VP1 and 3A proteins improve binding and replication of rhinovirus C15 in HeLa-E8 cells.
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.
['Bochkov|Yury A|YA|', 'Watters|Kelly|K|', 'Basnet|Sarmila|S|', 'Sijapati|Shakher|S|', 'Hill|Marchel|M|', 'Palmenberg|Ann C|AC|', 'Gern|James E|JE|']
D020816:Amino Acid Motifs D006367:HeLa Cells D006801:Humans D020125:Mutation, Missense D010850:Picornaviridae Infections D012229:Rhinovirus D014764:Viral Proteins D053585:Virus Attachment D014779:Virus Replication
Rhinovirus C Adaptation Heparan sulfate Pathogenesis Receptor specificity