Influence of signaling-pathway inhibitors on the T-cell stimulatory capacity of mature MDDC.
Maturation of MDDC was induced with TNF-α (20 ng/mL; A) or LPS (10 ng/mL; B,C) and in either the absence or presence of PD98059 (PD; 40 μM), U0126 (U0; 2.5 μM), or SB203580 (SB; 13 μM). After 48 hours, MDDC were irradiated and used to stimulate 2 × 105allogeneic peripheral blood T lymphocytes (A,B) or CD4+cord-blood lymphocytes (C) in 96-well plates. Stimulation was done at a 1:40 ratio (A) or the indicated ratios of MDDC to T cells (B,C). After a 5-day coculture, tritium-thymidine was added to the culture for 16 hours and T-cell proliferation determined by measuring the incorporated thymidine. Each experiment was done twice; representative experiments are shown.
