Fig. 3. In vitro dendritic cell (DC)–induced proliferation of allogeneic T cells and Th1 response in the presence (and absence) of midazolam. DCs were incubated with (and without) 15 μM midazolam and exposed to lipopolysaccharide for 24 h. DCs were treated with mitomycin C, then washed and cocultured with T cells derived from Balb/c mice. Results are presented as mean ± SD (N = 8). (A ) Mixed cell culture was performed for 4 days. Cell proliferation was estimated based on uptake of [3H]thymidine, which was pulsed during the final 18 h of culture. Radioactivity of harvested cells was measured using a liquid scintillation counter. Squares (▪) indicate DC treatment with phosphate buffer saline (PBS); circles (•) indicate treatment with midazolam. *P < 0.05, PBS-DC versus midazolam-DC. (B ) After 64 h of mixed cell culture, cell clustering was counted under microscopy. *P < 0.05, control versus midazolam.