TABLE 2.
Comparison of Core-NS2 SA13 (5a) JFH1-based recombinants in single-cycle virus production assayse
| Posttransfection time and virus | Intracellular infectivity titera (log10 FFU/well) | Extracellular infectivity titerb (log10 FFU/ml) | Ratio of extracellular/intracellular infectivity | Extracellular corec (log10 amol Core/ml) | Specific infectivity, extracellular (FFU/amol Core) |
|---|---|---|---|---|---|
| 24 h | |||||
| SA13/JFH1orig | 3.3 | 4.2 | 8 | 2.6 | 1/0.03 |
| SA13/JFH1orig,NS2 | 3.6 | 4.6 | 10 | 3.0 | 1/0.03 |
| SA13/JFH1p7-NS5B | 4.9 | 5.7 | 6 | 3.7 | 1/0.01 |
| SA13/JFH1Core-NS5B | 5.3 | 6.2 | 8 | 3.3 | 1/0.001 |
| J6/JFH1 | 3.8 | 4.7 | 8 | 3.6 | 1/0.08 |
| J6/JFH1-GNDd | NA | NA | NA | 1.8 | NA |
| 36 h | |||||
| SA13/JFH1orig | 4.7 | 5.9 | 16 | 4.5 | 1/0.04 |
| SA13/JFH1orig,NS2 | 4.9 | 5.9 | 10 | 4.6 | 1/0.05 |
| SA13/JFH1p7-NS5B | 5.8 | 6.7 | 8 | 5.0 | 1/0.02 |
| SA13/JFH1Core-NS5B | 6.4 | 7.0 | 4 | 4.7 | 1/0.005 |
| J6/JFH1 | 4.7 | 5.6 | 8 | 4.6 | 1/0.1 |
| J6/JFH1-GNDd | NA | NA | NA | 1.8 | NA |
| 48 h | |||||
| SA13/JFH1orig | 4.8 | 6.0 | 16 | 5.3 | 1/0.2 |
| SA13/JFH1orig,NS2 | 4.9 | 6.0 | 13 | 5.3 | 1/0.2 |
| SA13/JFH1p7-NS5B | 6.3 | 6.7 | 3 | 5.7 | 1/0.1 |
| SA13/JFH1Core-NS5B | 6.4 | 7.1 | 5 | 5.2 | 1/0.01 |
| J6/JFH1 | 4.7 | 4.9 | 2 | 5.0 | 1/1.3 |
| J6/JFH1-GNDd | NA | NA | NA | 1.8 | NA |
Intracellular infectivity titers were determined as described in Materials and Methods.
Extracellular infectivity titers were determined as described in Materials and Methods.
HCV Core titers were determined using the Architect HCV Ag assay (Abbott) and displayed as attomoles (amol) per milliliter.
For the J6/JFH1-GND negative control virus, infectivity titers were below cutoff (NA).
S29 cells (32) were transfected with RNA transcripts as described in the Fig. 3 legend and Materials and Methods. Supernatants and cell lysates were collected at 12, 24, 36, and 48 h posttransfection, and infectivity titers and HCV Core titers were determined. Only data from samples collected at 24, 36, and 48 h posttransfection are shown in the table since most data collected at 12 h posttransfection were below the cutoff.