TABLE 2.
Markers of apoptosis in DC inoculated with wt, ΔNS1, ΔNS2, or ΔNS1/2 RSVa
| Virus | % Annexin V positive
|
% Active caspase 3 positive
|
DNA fragmentation ELISA (optical density)
|
|||||||
|---|---|---|---|---|---|---|---|---|---|---|
| GFP-positive DC
|
Total DC
|
GFP-positive DC
|
Total DC
|
Total DC
|
||||||
| 24 h | 48 h | 24 h | 48 h | 24 h | 48 h | 24 h | 48 h | 24 h | 48 h | |
| wt RSV | 2.3 | 65.1 | 0.6 | 3.2 | 4.4 | 4.0 | 1.2 | 6.3 | 0.1 | 0.5 |
| ΔNS1 | 3.8 | 39.4 | 0.9 | 2.5 | 3.6 | 1.7 | 0.8 | 2.8 | 0.3 | 0.3 |
| ΔNS2 | 7.0 | 65.9 | 1.0 | 5.0 | 8.9 | 3.5 | 1.2 | 6.6 | 0.3 | 0.3 |
| ΔNS1/2 | 2.6 | 51.7 | 0.6 | 3.6 | 1.5 | 1.8 | 0.5 | 2.8 | 0.3 | 0.3 |
| Mock | NAb | NA | 1.4 | 16 | NA | NA | 6.4 | 24.7 | 1.0 | 1.0 |
| UV-wt | NA | NA | 1.7 | 8.2 | NA | NA | 6.6 | 10.4 | 0.7 | 0.6 |
| Positive controlc | NA | NA | 14 | 30 | NA | NA | 32.3 | 78 | 2.8 | 2.6 |
Immature DC were inoculated with the indicated viruses at an input MOI of 2 PFU/cell, and percentages of annexin V-positive or caspase-3-positive cells were determined for both the total DC and GFP-positive DC populations at 24 and 48 h postinfection. The extent of DNA fragmentation was determined for the total population at 24 and 48 h postinfection by ELISA. Three donors are represented: each assay involved a different donor.
NA, not applicable, since no green fluorescent cells were detected in the samples.
The positive control in the annexin V and activated caspase-3 assays was immature DC treated with 5 μM staurosporine; the positive control for the DNA fragmentation ELISA was provided with the kit.