A New Model to Produce Infectious Hepatitis C Virus without the Replication Requirement
Figure 6
JFH-1-based HCV produced in BHK-WNV cells infect Huh-7.5 cells.
Huh-7.5 cells were seeded one day before the experiment and treated with (open symbols, dashed lines) or without (closed symbols) leukocyte interferon (100 IU/ml) plus ribavirin (400 µM). JFH-1 (A) and Con1-JFH1 (B) HCV particles produced in BHK-WNV cells were incubated with the Huh-7.5 cells for 2 hrs; after several washes, the cells were split and an equivalent amount of cells were either directly harvested (day 0) or seeded onto collagen-I-coated 24-well plates (and interferon plus ribavirin treatment was re-introduced where applies) and further cultured for the indicated times. Cells were then harvested and HCV 5′-UTR RNA was subject to RT-qPCR analysis, as in (Fig. 4A). Errors bars represent SD of 4 measurements; the limit of detection in this assay is indicated by a dotted line; nd = undetermined values (i.e. <105 HCV RNA copies/well).