Interaction of immunosuppressants with HCV antivirals daclatasvir and asunaprevir: combined effects with mycophenolic acid

Figure 3 Calcineurin inhibitor cyclosporine A does not affect the antiviral activity of direct acting antivirals. A, B: Huh7-ETluc cells were cultured with increasing concentrations of DCV (A) or ASV (B), in combination with different concentrations of CSA. After 24 h luciferase was measured. HCV replication was inhibited by CSA: 5 μg/mL CSA significantly inhibited HCV replication compared to control (Mann-Whitney test, P = 0.03 for DCV, P = 0.04 for ASV). The antiviral action of DCV of ASV was not negatively affected by CSA and vice versa; C, D: The luciferase signal in Huh7-PGK-luc cells, stably expressing luciferase, was not affected by any concentration of DCV (C) or ASV (D) with or without CSA, indicating that the observed effect in HUh7-ETluc is not due to non-specific inhibition of luciferase. Results are mean ± SE of 4 independent experiments performed in triplicate; E, F: In the JFH infectious HCV cell culture model, HCV RNA levels were inhibited to by > 99% of control levels by both DCV (E) and ASV (F). The addition of 0.5 μg/mL CSA completely inhibited HCV replication (E, F). Shown are the results of two independent experiments, measured in duplicate by RT-qPCR. CSA: Cyclosporine A; HCV: Hepatitis C virus; DCV: Daclatasvir; ASV: Asunaprevir.