Table 3.
PMA/ionomycin and LPS-stimulated Mφ IL-10 and TNF-α are PKA independent
| IL-10 pg/ml (% control) | TNF-α pg/ml (% control) | |
|---|---|---|
| PMA/ionomycin | 46·77 ± 1·285 (100) | 3303 ± 207·4 (100) |
| + H89 10 µm | 44·73 ± 2·236 (96) | 3079 ± 25·32 (93) |
| 100 µm | 45·36 ± 2·445 (97) | 3312 ± 40·22 (100) |
| 1000 µm | 41·57 ± 2·950 (89) | 2690 ± 207·4 (81) |
| LPS | 1946 ± 57·81 (100) | 1296 ± 39·06 (100) |
| + H89 10 µm | 2085 ± 81·75 (107) | 1265 ± 126·4 (98) |
| 100 µm | 1912 ± 223·6 (98) | 1357 ± 25·90 (105) |
| 1000 µm | 1681 ± 318·0 (86) | 1288 ± 344·4 (99) |
Monocytes primed with M-CSF for 7 days (Mφs) were plated at a density of 1 × 105 cells per well in 96-well flat bottomed plates and were preincubated for 1 hr with the PKA inhibitor, H89 or the appropriate vehicle control, afterwhich cells were stimulated with PMA/ionomycin (50 ng/ml and 0·5 µg/ml, respectively) or LPS (1 ng/ml) and incubated for a further 24 hr. Supernatants were harvested and assayed for IL-10 and TNF-αlevels by ELISA. Data are mean cytokine levels in pg/ml of triplicate culture supernatants ± SD and their percentage change over control levels (100%), showing a representative of two replicate experiments.