FIG 7.

Antiviral activity of the kinase inhibitor EMD 1214063 (EMD). (A) Inhibition of different viruses with EMD 1214063. Monolayers of A549 cells were pretreated with the indicated concentration of EMD 1214063 for 30 min, followed by infection with the different viruses (MOI = 0.3) in the presence of drug. After 1 h, the cells were washed with medium containing 20 mM ammonium chloride. Infection of rLCMV-LASVGP and LCMV was detected after 16 h by IFA as described in the legend to Fig. 2B. IAV infection was assessed after 6 h by staining for IAV N protein, whereas AdV was detected via its EGFP reporter by direct fluorescence after 24 h. Data are means ± SDs (n = 3). (B) Representative images of inhibition experiments. LCMV and IAV antigens are in red, EGFP is in green, and nuclei are stained with DAPI (blue). Bar = 100 μM. (C) Antiviral effect of EMD 1214063 in primary human respiratory epithelial cells. Monolayers of primary human bronchiolar epithelial cells isolated from human bronchi were cultured in M96 plates for 3 days to obtain closed cell monolayers. The cells were pretreated with the indicated concentration of EMD 1214063 for 30 min, followed by infection with rLCMV-LASVGP (LASV) and rLCMV-VSVG (VSV) at 1,000 PFU/well for 1 h. The cells were washed, and infection was detected by IFA after 16 h, as described in the legend to Fig. 2B. Data are means ± SDs (n = 3).