title: Development of a high-throughput human rhinovirus infectivity cell-based assay for identifying antiviral compounds.
abstract: Asthma and chronic obstructive pulmonary disease exacerbations are associated with human rhinovirus (HRV) lung infections for which there are no current effective antiviral therapies. To date, HRV infectivity of cells in vitro has been measured by a variety of biochemical and immunological methods. This paper describes the development of a high-throughput HRV infectivity assay using HeLa OHIO cells and a chemiluminescent-based ATP cell viability system, CellTiter-Glo from Promega, to measure HRV-induced cytopathic effect (CPE). This CellTiter-Glo assay was validated with standard antiviral agents and employed to screen AstraZeneca compounds for potential antiviral activity. Compound potency values in this assay correlated well with the quantitative RT-PCR assay measuring HRV infectivity and replication in human primary airway epithelial cells. In order to improve pan-HRV screening capability, compound potency was also measured in the CellTiter-Glo assay with a combination of 3 different HRV serotypes. This HRV serotype combination assay could be used to identify quickly compounds with desirable broad spectrum antiviral activity.
authors.affiliation: Bioscience, AstraZeneca R&D Charnwood, Loughborough, Bakewell Road, Leicestershire LE11 5RH, United Kingdom. tim.phillips@astrazeneca.com
authors.forename: Tim
authors.initials: T
authors.lastname: Phillips
authors.forename: Lesley
authors.initials: L
authors.lastname: Jenkinson
authors.forename: Christopher
authors.initials: C
authors.lastname: McCrae
authors.forename: Bob
authors.initials: B
authors.lastname: Thong
authors.forename: John
authors.initials: J
authors.lastname: Unitt
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods; D008826:Microbial Sensitivity Tests / Q000379:methods; D012229:Rhinovirus / Q000187:drug effects* / Q000254:growth & development; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents