Persistent Growth of a Human Plasma-Derived Hepatitis C Virus Genotype 1b Isolate in Cell Culture
Figure 4
Inhibition of HCV by antivirals.
(A) LB-piVe cells (B) and J6/JFH1-infected Huh7.5 cells were treated with increasing concentrations of 2′-C-Me-A. The EC50 values were evaluated from dose response curves employing GraphPad Prism 3.0 software. (C) Non-specific siRNA (IRR) [64] or HCV-specific siRNA (313) [38] transfected Huh7.5 cells were inoculated with LB-piVe and stained with crystal violet (left). Filter-clarified culture supernatants were titrated by a CPE-based end-point dilution assay (right). (D) Non-specific siRNA (IRR) [64] or HCV-specific siRNA (313) [38] transfected Huh7.5 cells were inoculated with J6/JFH-1 and stained with anti-Core antibodies at 3 dpi (left). Fluorescent foci were counted in triplicate wells, and titers were calculated as the mean number of foci per ml (FFU/ml, right). (E) LB-piVe cells were transfected with pVA and then treated with 0, 10, 100 and 1000 IU/ml of Universal Type 1 IFN for 24, 48 and 72 hr. Viral titers were determined as in B. (F) Huh7.5 cells were inoculated with J6/JFH-1 and treated with 0, 10, and 1000 IU/ml of Universal Type 1 IFN for 72 hr. Viral titers were determined by infecting Huh7.5 cells in the absence of pVA. (G) LB-piVe titers in the absence of pVA, determined in naïve Huh7.5 cells with (+IFN) or without (−IFN) the addition of 0, 10, and 1000 IU/ml of Universal Type I IFN to the culture media. (H) Infection of naïve, non-transfected VeroE6 cells with genotype 1b-infectious human plasma (LB; [28], [29]) and treated with 1000 IU/mL IFN. HCV RNA copies were determined per µg of GAPDH RNA. HCV titers were calculated using the method of Reed and Muench [31]. Error bars, ±SD.