TABLE 1.
Replication of rDIsSIVGag and other vaccinia virus recombinants and production of SIV Gag p27 proteina
| Cell type and vaccinia virus | Mean amt of virus replicationb ± SD | Mean p27 antigen amt (pg/ml) ± SD in 106 cells
|
|
|---|---|---|---|
| Supernatant | Cell lysate | ||
| CEF cells | |||
| rWRSIVGag | 9,500 ± 1,800 | 6,530 ± 2,600 | 87,300 ± 14,800 |
| rDIsSIVGag | 1,860 ± 350 | 2,470 ± 60 | 15,800 ± 3,700 |
| rDIsLacZ | 1,400 ± 240 | <20 | <20 |
| MVA | 10,327 ± 1,479 | <20 | <20 |
| BHK-21 cells | |||
| rWRSIVGag | 2,500 ± 400 | 970 ± 22 | 34,600 ± 12,700 |
| rDIsSIVGag | <1 | <20 | 1,375 ± 273 |
| rDIsLacZ | <1 | <20 | <20 |
| MVA | 2,250 ± 835 | <20 | <20 |
| RK13 cells | |||
| rWRSIVGag | 1,500 ± 300 | 350 ± 12 | 16,300 ± 9,270 |
| rDIsSIVGag | <1 | <20 | 275 ± 34 |
| rDIsLacZ | <1 | <20 | <20 |
| MVA | 125 ± 102 | <20 | <20 |
| CHO cells | |||
| rWRSIVGag | 953 ± 347 | 410 ± 120 | 5,320 ± 7,670 |
| rDIsSIVGag | <1 | <20 | 119 ± 34 |
| rDIsLacZ | <1 | <20 | <20 |
| MVA | 92 ± 49 | <20 | <20 |
CEF, RK13, and BHK-21 cells were infected with rDIs or rWR strain viruses at an MOI of 0.05 for the detection of virus replication and at an MOI of 1.0 for p27 antigen generation in the culture supernatant and cells. p27 antigen was measured by enzyme-linked immunosorbent assay in cultures containing 106 cells/ml, and the data are presented as the average value ± the SD.
For analysis of virus replication, mammalian cells were infected with each virus at an MOI of 0.05. The cells were harvested at 0 and 48 h after adsorption and sonicated. Virus replication was determined by dividing the virus yield at 48 h by that at 0 h (15). Values of <1 and <20 pg/ml represent the detection limits of the virus replication and p27 antigen assays, respectively.