Ifn alpha α

Vero cells were grown in 24-well plates at a density of 50,000 cells/cm², the day before infection. First, the cells were treated with human interferon (IFN) alpha (α; PBL Assay Science, Piscataway, NJ, USA) and beta (β; R&D Systems, Mineapolis, MI, USA) at concentrations of 10 UI/mL, 50 UI/mL, 100 UI/mL, and 1000 UI/mL for 6 h before infection. Next, cell supernatants were removed, and 100 µL of viral suspensions were added at MOI of 0.5. After 1 h incubation, the viral inocula were discarded, and 0.3 mL of supplemented Earle’s 199 medium containing the four different IFN concentrations were added to the cells. After 48 h of incubation, the cell supernatants were collected for viral titration via plaque assay.
The viral titers under treatment with IFN-α and IFN-β were normalized with the values obtained from non-treated and infected cells. This experiment was performed in three independent assays for statistical relevance. Data were analyzed in Prism version 8 (GraphPad Software, San Diego, CA, USA). IC₅₀ values were calculated from the nonlinear regression function provided by the software ([Inhibitor] vs. normalized response—Variable slope). IC₅₀ values of each replicate were analyzed using a one-way ANOVA with Dunnett’s multiple comparisons test.

Vero cells were grown in 24-well plates at a density of 50,000 cells/cm². The next day, cells were treated with interferon (IFN) alpha (α; PBL Assay Science, United States) and beta (β; R&D Systems, United States) at concentrations of 10, 50, 100, and 1,000 UI/mL for 6 h before infection. Cell supernatant was removed, and 100 μL of viral suspensions were added at MOI of 0.5. After 1 h incubation with gentle agitation every 15 min, viral inocula were discarded, and cells were overlaid with 0.3 mL of supplemented Earle’s 199 medium, containing the four different IFN concentrations. After 48 h incubation at 37°C and 5% CO₂, the supernatants were collected for viral titration by plaque assay. The log₁₀ of viral titers under treatment with IFN-α and IFN-β were normalized with the values obtained from non-treated infected cells. This experiment was performed in three independent assays for statistical relevance. Data were analyzed in GraphPad Prism 8 software. IC₅₀ values were calculated from the non-linear regression function provided by the software [(Inhibitor) vs. normalized response – Variable slope]. IC₅₀ values of each replicate were analyzed by one-way ANOVA with Bonferroni’s multiple comparison test.