TABLE 1.
Involvement of TNF-α in the suppressive activity of LNCi
| Culturea | Proliferation (cpm)b | NO production (μM)c | IFN-γ production (U/ml)d |
|---|---|---|---|
| LNCn (WT) | 220,952 ± 1,752 | 2.5 ± 1 | 4 ± 0.7 |
| LNCn (TNF-α−/−) | 226,699 ± 3,111 | 2.2 ± 0.8 | 3.1 ± 0.8 |
| LNCn (WT) + LNCi (WT) | 831 ± 121 | 44.5 ± 5.0 | 511 ± 42 |
| LNCn (WT) + LNCi (TNF-α−/−) | 203,378 ± 18,740 | 31.5 ± 4.3 | 138 ± 11 |
LNC cultures at a concentration of 2 × 106/ml were stimulated with ConA. Cocultures of LNCn (2 × 106/ml) and LNCi (2 × 106/ml) were made in order to evaluate the suppressive activity of LNCi on the response of LNCn. The role of TNF-α in the suppressive activity was analyzed by using LNCi from wild-type (WT) mice as well as LNCi from TNF-α−/− mice.
Proliferation of cell cultures was measured by [3H]thymidine incorporation after 48 h of incubation in the presence of ConA. Representative results are shown as mean cpm ± standard deviation.
NO production was quantified by nitrite accumulation (Griess reaction) in the supernatants of cell cultures stimulated with ConA for 48 h. Results from triplicate measurements are shown as means ± standard deviations.
IFN-γ production was quantified by specific ELISA (Pharmingen) in the supernatants of cell cultures stimulated with ConA for 48 h. Results from triplicate measurements are shown as means ± standard deviations.