| Assay Method Information | |
| | Cellular Assay-Influenza Virus Replication PR8 and X31 in Calu-3 Human Bronchial Epithelial Cells |
| Description: | The ability of the tested compound to block influenza virus replication (PR8 and X31) in Calu-3 human bronchial epithelial cells was evaluated as described by Beaulieu A. et al. J Virol. 2013 April; 87(8):4237-51.Calu-3 cells were washed with Dulbecco's phosphate-buffered saline (D-PBS) and exposed to influenza virus (diluted in incomplete medium; 0.2% bovine serum albumin [BSA] instead of FBS). After virus adsorption (1 h at 37° C.), cells were washed once with D-PBS, and cells were incubated in incomplete culture medium containing increasing concentrations of the tested compound for 48 h.Viral titers were determined in the supernatants of infected cells by viral plaque assays as described by Cloutier et al. J Infect Dis. 2012 Feb. 15; 205(4):621-30. Serial 10-fold dilutions of clarified supernatants were prepared in incomplete Eagle's minimal essential medium (EMEM) (containing 0.1% bovine serum albumin instead of fetal bovine serum) and were titered on Madin-Darby canine kidney (MDCK) cells according to standard viral plaque assays. Confluent MDCK cells were exposed to lung supernatant dilutions for 1 hour to allow virus adsorption. Cells were then washed, and a semifluid medium containing Avicel RC-581 (FMC BioPolymer), incomplete EMEM, and 1 μg/mL Tosyl phenylalanyl chloromethyl ketone (TPCK)-treated trypsin (Sigma-Aldrich) was added to the cells. Cells were incubated for 48 hours, and viral plaques were revealed with 2% crystal violet after Carnoy fixation. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |