Table 2.
Characteristics of HCV DBN3acc mutants after transfection and subsequent passage in Huh7.5 cells.
| Mutant | Spread (day)a | Infectivity titre (day)b | Fold Change EC50c | Engineered mutations |
|---|---|---|---|---|
| DBN3acc | 10 | 4.65 (10) | N/A | Maintained |
| NS2 119A | 10 | 4.71 (10) | 1.02 | Maintained |
| NS2 119V | 10 | 4.76 (10) | 1.22 | Maintained |
| NS2 132V | 17 | 4.10 (23) | 0.61 | Maintainedd |
| NS3 67A | 10 | 4.49 (10) | 0.65 | Maintained |
| NS5B 150V | 10 | 4.50 (14) | 1.00 | Maintained |
| NS2 119A + NS2 132V | 10 | 4.62 (14) | 1.14 | Maintained |
| NS2 119V + NS2 132V | 10 | 4.33 (14) | 0.60 | Maintained |
| NS2 119A + NS3 67A | 10 | 4.51 (10) | 1.19 | Maintained |
| NS2 119V + NS3 67A | 10 | 4.40 (10) | 1.06 | Maintained |
| NS2 119V + NS2 132V+ NS3 67A | 10 | 4.56 (14) | 0.98 | Maintained |
| NS2 119A + NS2 132V + NS5B 150Ve | N/A | N/A | N/A | N/A |
| NS2 119V + NS2 132V + NS5B 150V | 10 | 4.39 (14) | 0.98 | Maintained |
| NS2 119A + NS3 67A + NS5B 150V | 10 | 4.14 (14) | 0.79 | Maintained |
Viral kinetics, sofosbuvir susceptibility and genetic stability of DBN3acc original and mutant viruses.
aDay of spread after transfection, which corresponds to the day where ≥80% HCV antigen-positive cells were observed in the culture.
bThe highest infectivity titre observed in supernatants harvested after transfection with the day indicated in parenthesis. Data is based on the mean of three replicates and the titre units are Log10 focus-forming units per millilitre (FFU/ml).
cChange in sofosbuvir susceptibility shown as fold-change in EC50 values between original and mutant viruses.
dThe DBN3acc-NS2I132V mutant virus acquired substitutions A4506A/G (aa I1389I/M in NS3) and A7480A/G (aa T2381T/A in NS5A) as detected during passage.
eRecombinant DBN3acc- NS2 119A + NS2 132V + NS5B 150V was non-viable in two independent transfections. N/A: not applicable.