Effect of Incubation Time and Neutrophil Presence on the Concentration of TNF-α in Supernatants
| Time . | Medium + TNF-α . | Neutrophils + TNF-α . | Neutrophils − TNF-α . |
|---|---|---|---|
| (h) . | (ng/mL) . | (ng/mL) . | (ng/mL) . |
| 0 | 8.87 ± 0.59 | 10.29 ± 1.10 | 0* |
| 3 | 8.33 ± 0.54 | 7.84 ± 0.62 | 0 |
| 6 | 9.18 ± 0.90 | 8.67 ± 0.97 | 0 |
| 12 | 8.96 ± 1.08 | 8.29 ± 0.68 | 0 |
| 20 | 8.69 ± 0.63 | 7.97 ± 0.67 | 0 |
| Time . | Medium + TNF-α . | Neutrophils + TNF-α . | Neutrophils − TNF-α . |
|---|---|---|---|
| (h) . | (ng/mL) . | (ng/mL) . | (ng/mL) . |
| 0 | 8.87 ± 0.59 | 10.29 ± 1.10 | 0* |
| 3 | 8.33 ± 0.54 | 7.84 ± 0.62 | 0 |
| 6 | 9.18 ± 0.90 | 8.67 ± 0.97 | 0 |
| 12 | 8.96 ± 1.08 | 8.29 ± 0.68 | 0 |
| 20 | 8.69 ± 0.63 | 7.97 ± 0.67 | 0 |
Human neutrophils (5 × 106/mL) or medium alone (MDM containing 10% autologous serum) were incubated in the presence (+TNF-α) or absence (−TNF-α) of 12.5 ng/mL TNF-α in flexiwell plates in an humidified 5% CO2 incubator at 37°C for 0 to 20 hours. The supernatants were harvested, frozen, and immunoreactive human TNF-α quantified by ELISA as detailed in the Materials and Methods section. Data represent mean ± SEM of six measurements performed in two separate experiments.
Assay sensitivity 15.6 pg/mL.