Effect of rAAV Infection on Proliferation and Immunophenotype of Cells Developing in the T-Lymphopoiesis System
| . | . | Expansion Index . | Immunophenotype . | |||
|---|---|---|---|---|---|---|
| . | . | . | CD2+ . | CD3+ . | CD4+ . | CD8+ . |
| Day 21 | Control | 11.6 ± 4.4 | 29.2 ± 2.0 | 18.9 ± 5.5 | 51.8 ± 12.7 | 9.8 ± 2.5 |
| +rAAV | 12.4 ± 3.6 | 24.7 ± 5.4 | 16.7 ± 5.2 | 50.7 ± 11.2 | 10.8 ± 2.5 | |
| Day 35 | Control | 86.3 ± 65.4 | 73.5 ± 5.8 | 89.1 ± 4.7 | 87.4 ± 4.3 | 10.5 ± 3.7 |
| +rAAV | 82.0 ± 53.7 | 68.7 ± 4.3 | 85.4 ± 6.1 | 83.9 ± 3.9 | 14.0 ± 5.3 | |
| . | . | Expansion Index . | Immunophenotype . | |||
|---|---|---|---|---|---|---|
| . | . | . | CD2+ . | CD3+ . | CD4+ . | CD8+ . |
| Day 21 | Control | 11.6 ± 4.4 | 29.2 ± 2.0 | 18.9 ± 5.5 | 51.8 ± 12.7 | 9.8 ± 2.5 |
| +rAAV | 12.4 ± 3.6 | 24.7 ± 5.4 | 16.7 ± 5.2 | 50.7 ± 11.2 | 10.8 ± 2.5 | |
| Day 35 | Control | 86.3 ± 65.4 | 73.5 ± 5.8 | 89.1 ± 4.7 | 87.4 ± 4.3 | 10.5 ± 3.7 |
| +rAAV | 82.0 ± 53.7 | 68.7 ± 4.3 | 85.4 ± 6.1 | 83.9 ± 3.9 | 14.0 ± 5.3 | |
The expansion index was determined by dividing the number of viable cells present at different time points by the number of starting CD34+CD2− cells from independent experiments (n = 6). Trypan blue staining was used to evaluate cell viability, which was routinely greater than 90%. Immunophenotype was evaluated for a subset of the experiments (n = 3) by fluorescent antibody staining and FACS-quadrant analysis as described for Fig 2. Total percentages of cells expressing individual antigens are shown, and the dominant cell population present at day 21 was CD2+CD3+CD4+, whereas at day 35, maturation to single-positive CD3+CD4+ and CD3+CD8+ T cells had occurred. All data are the mean ± SD from duplicate readings.