Relative gene transfer efficiencies and long-term transgene expression in mitogen-stimulated and resting primary human lymphocytes transduced with HIV-1-based vectors
| Cell sample . | . | APs . | Percentage of EGFP-positive cells after culture . | ||||
|---|---|---|---|---|---|---|---|
| 3 days . | 2 wk . | 4 wk . | 8 wk . | 16 wk . | |||
| PBLs* | Stimulated | + | 96.4 ± 1.0 | 70.6 ± 1.8 | 53.8 ± 1.8 | 38.8 ± 2.4 | 26.7 ± 2.9† |
| Δ | 93.0 ± 1.3 | 71.2 ± 0.6 | 45.6 ± 2.0 | 33.0 ± 1.7 | 25.3 ± 2.7† | ||
| Resting | + | 90.2 ± 1.1 | 69.0 ± 3.2 | 49.8 ± 1.3 | 34.6 ± 1.9 | 22.3 ± 0.7† | |
| Δ | 4.8 ± 0.7 | 3.6 ± 0.7 | 3.2 ± 0.4 | 2.5 ± 0.4 | 2.3 ± 0.3† | ||
| CBLs‡ | Stimulated | + | 89.0 ± 2.0 | 71.0 ± 3.0 | 53.0 ± 4.5 | 35.0 ± 0.1 | 29.02-153 |
| Δ | 89.0 ± 1.5 | 71.0 ± 0.5 | 43.0 ± 5.0 | 26.0 ± 1.5 | 23.02-153 | ||
| Resting | + | 86.0 ± 6.5 | 65.0 ± 4.0 | 43.0 ± 7.0 | 33.0 ± 6.0 | 18.02-153 | |
| Δ | 5.0 ± 2.0 | 4.0 ± 2.0 | 3.5 ± 1.5 | 2.5 ± 1.5 | ND | ||
| Cell sample . | . | APs . | Percentage of EGFP-positive cells after culture . | ||||
|---|---|---|---|---|---|---|---|
| 3 days . | 2 wk . | 4 wk . | 8 wk . | 16 wk . | |||
| PBLs* | Stimulated | + | 96.4 ± 1.0 | 70.6 ± 1.8 | 53.8 ± 1.8 | 38.8 ± 2.4 | 26.7 ± 2.9† |
| Δ | 93.0 ± 1.3 | 71.2 ± 0.6 | 45.6 ± 2.0 | 33.0 ± 1.7 | 25.3 ± 2.7† | ||
| Resting | + | 90.2 ± 1.1 | 69.0 ± 3.2 | 49.8 ± 1.3 | 34.6 ± 1.9 | 22.3 ± 0.7† | |
| Δ | 4.8 ± 0.7 | 3.6 ± 0.7 | 3.2 ± 0.4 | 2.5 ± 0.4 | 2.3 ± 0.3† | ||
| CBLs‡ | Stimulated | + | 89.0 ± 2.0 | 71.0 ± 3.0 | 53.0 ± 4.5 | 35.0 ± 0.1 | 29.02-153 |
| Δ | 89.0 ± 1.5 | 71.0 ± 0.5 | 43.0 ± 5.0 | 26.0 ± 1.5 | 23.02-153 | ||
| Resting | + | 86.0 ± 6.5 | 65.0 ± 4.0 | 43.0 ± 7.0 | 33.0 ± 6.0 | 18.02-153 | |
| Δ | 5.0 ± 2.0 | 4.0 ± 2.0 | 3.5 ± 1.5 | 2.5 ± 1.5 | ND | ||
Primary human lymphocytes isolated from PB and CB samples were either left unstimulated or stimulated with PHA and rhIL-2 for 3 days and then transduced with VSV-G pseudotyped pHR′PGK-EGFP viral supernatants generated in the presence (+) or the absence (Δ) of HIV-1 accessory proteins (APs) at an MOI of 5 to 10. After transduction, the resting lymphocytes were stimulated with mitogens and cultured in vitro in medium containing rhIL-2 and periodically stimulated with PHA. The number of EGFP-positive cells (mean ± SE) was determined at indicated time points by FACS analysis.
Values obtained from 5 samples unless otherwise noted.
Values obtained from 3 samples.
Values obtained from 2 samples, unless otherwise noted.
Data obtained from one sample.
ND = not done.