Dexamethasone Suppressed Human IL-5 Promoter/Enhancer Activity Induced by Either TCR or IL-2R Stimulation
| Clone* . | Stimulation† . | Dex‡ . | Luciferase Activityρ (per μg protein) . | β-Galactosidase Activityρ (per μg protein) . |
|---|---|---|---|---|
| YA5 | None | − | 10.9 ± 1.0 | 0.67 ± 0.07 |
| Anti-CD3 | − | 192.4 ± 12.2 | 0.59 ± 0.08 | |
| + | 21.0 ± 2.0 | 0.62 ± 0.07 | ||
| IL-2 | − | 118.2 ± 10.0 | 0.66 ± 0.08 | |
| + | 17.2 ± 1.2 | 0.69 ± 0.06 | ||
| HK5 | None | − | 4.2 ± 1.0 | 0.92 ± 0.08 |
| Anti-CD3 | − | 479.2 ± 30.9 | 0.87 ± 0.10 | |
| + | 12.3 ± 1.0 | 0.97 ± 0.13 | ||
| HK2 | None | − | 8.2 ± 1.0 | 0.72 ± 0.08 |
| IL-2 | − | 269.2 ± 20.1 | 0.67 ± 0.11 | |
| + | 9.2 ± 2.9 | 0.77 ± 0.10 |
| Clone* . | Stimulation† . | Dex‡ . | Luciferase Activityρ (per μg protein) . | β-Galactosidase Activityρ (per μg protein) . |
|---|---|---|---|---|
| YA5 | None | − | 10.9 ± 1.0 | 0.67 ± 0.07 |
| Anti-CD3 | − | 192.4 ± 12.2 | 0.59 ± 0.08 | |
| + | 21.0 ± 2.0 | 0.62 ± 0.07 | ||
| IL-2 | − | 118.2 ± 10.0 | 0.66 ± 0.08 | |
| + | 17.2 ± 1.2 | 0.69 ± 0.06 | ||
| HK5 | None | − | 4.2 ± 1.0 | 0.92 ± 0.08 |
| Anti-CD3 | − | 479.2 ± 30.9 | 0.87 ± 0.10 | |
| + | 12.3 ± 1.0 | 0.97 ± 0.13 | ||
| HK2 | None | − | 8.2 ± 1.0 | 0.72 ± 0.08 |
| IL-2 | − | 269.2 ± 20.1 | 0.67 ± 0.11 | |
| + | 9.2 ± 2.9 | 0.77 ± 0.10 |
IL-5–producing T-cell clones (2.5 × 107 cells in 500 mL RPMI1640 medium supplemented with 10% FBS and 10 U/mL rIL-2) were transiently transfected with pIL-5Luc (50 μg) + pCMV-β-gal (10 μg) by electroporation (270 V, 960 μF) using a 0.4-cm cuvette. Transfected cells were suspended in a fresh medium and cultured in triplicate in 24-well culture plates.
Cells were either stimulated with rIL-2 (100 U/mL) or immobilized anti-CD3 MoAb or kept unstimulated for 24 hours.
Dexamethasone (1 μmol/L) was included from the start of some cultures, as indicated..
ρ After 24 hours, cell lysates were prepared and tested for luciferase activity using Luciferase Assay System and β-galactosidase activity using CPRG as a substrate. Protein concentrations were determined with bicinchoninic acid protein assay reagent. Data are presented as the mean of triplicate cultures ± SEM.