Reversion of the Inhibitory Effect of RCC CM by Neutralizing Antibodies Against (IL-6 + IL-6R) or M-CSF
| Conditions . | Control . | CLB-VER CM (10%) . | IL-6 (20 ng/mL) (cpm × 10−3) . | M-CSF (20 ng/mL) . | IL-6 + M-CSF (20 ng/mL) . |
|---|---|---|---|---|---|
| Control antibodies | 29.7 ± 3.4 | 7.6 ± 1.4 | 12.9 ± 1.3 | 11.5 ± 1.4 | 11.9 ± 0.5 |
| Anti–(IL-6 + IL-6R) | 30.0 ± 2.9 | 13.0 ± 1.4 | 29.1 ± 3.0 | 17.9 ± 1.8 | 17.7 ± 3.0 |
| Anti–M-CSF | 29.6 ± 1.8 | 19.2 ± 3.4 | 15.7 ± 2.0 | 27.0 ± 2.4 | 14.8 ± 1.3 |
| Anti–(IL-6 + IL-6R) + anti–M-CSF | 29.9 ± 1.3 | 25.5 ± 1.5 | 32.1 ± 3.4 | 27.4 ± 1.1 | 25.0 ± 2.6 |
| Anti-VEGF | 30.5 ± 1.8 | 7.7 ± 0.9 | 13.1 ± 1.1 | 11.3 ± 0.7 | 12.3 ± 1.5 |
| Conditions . | Control . | CLB-VER CM (10%) . | IL-6 (20 ng/mL) (cpm × 10−3) . | M-CSF (20 ng/mL) . | IL-6 + M-CSF (20 ng/mL) . |
|---|---|---|---|---|---|
| Control antibodies | 29.7 ± 3.4 | 7.6 ± 1.4 | 12.9 ± 1.3 | 11.5 ± 1.4 | 11.9 ± 0.5 |
| Anti–(IL-6 + IL-6R) | 30.0 ± 2.9 | 13.0 ± 1.4 | 29.1 ± 3.0 | 17.9 ± 1.8 | 17.7 ± 3.0 |
| Anti–M-CSF | 29.6 ± 1.8 | 19.2 ± 3.4 | 15.7 ± 2.0 | 27.0 ± 2.4 | 14.8 ± 1.3 |
| Anti–(IL-6 + IL-6R) + anti–M-CSF | 29.9 ± 1.3 | 25.5 ± 1.5 | 32.1 ± 3.4 | 27.4 ± 1.1 | 25.0 ± 2.6 |
| Anti-VEGF | 30.5 ± 1.8 | 7.7 ± 0.9 | 13.1 ± 1.1 | 11.3 ± 0.7 | 12.3 ± 1.5 |
The CD34+ cells were cultured in presence of GM-CSF + TNFα alone (control) or associated with RCC CM, IL-6, M-CSF, or IL-6 + M-CSF in the presence of neutralizing antibodies against IL6 + IL-6R (10 μg/mL), M-CSF (10 μg/mL), VEGF (10 μg/mL), or control antibodies (10 μg/mL). Cultured CD34+ cells were harvested at day 12, irradiated (30 Gy), and used as stimulator cells (104 cells) in an allogeneic MLR with CD45RA+naive T cells (2 × 104 cells/well) as described in Materials and Methods. Results are expressed as the mean ± SD of triplicate culture. Results are representative of four experiments.