Anti-estrogens inhibit the terminal maturation of DC.
(A) Adherent PBMC were cultured for 7 days with IL-4 + GM-CSF in the presence or absence of TOR. On days 3 and 5, 200 μL fresh medium containing IL-4 (500 U/mL) + GM-CSF (50 ng/mL) was added with or without TOR (2.5 or 5 μmol/L). The cells were then cocultured with 2 × 105 CD40L-transfected J558L cells for 48 hours. The levels of biologically active IL-12 p70 were measured from culture supernatants by ELISA. The data represent mean cytokine levels ± SD of 3 individual experiments. The control J558L cells did not induce IL-12 production (not shown). (B) The cells were cultured as described in (A). A representative experiment of 3 experiments comparing TOR and TAM on the IL-12 p70 production after CD40 ligation is shown. (C) The immature DC were cultured in a medium alone (control) or activated with LPS (1 μg/mL) in the presence or absence of TOR or TAM. After 36 hours the cells were washed, irradiated (30 Gy), and counted. Five to 20 × 103 cells were then cocultured with 2 × 105 allogeneic PBMC for 5 days, after which the proliferative activity was measured. The results are expressed as mean cpm ± SD of a representative experiment made in triplicate. Similar results were obtained in 2 other experiments.
