Figure 2
Cigarette smoke extract and cigarette smoking promote dendritic cell survival. A) Human dendritic cells (1 × 106/ml in complete media) were incubated in the presence of absence of CSE (0.01 to 1%) for 24 hours prior to determination of cellular viability using the XTT assay. *p < 0.05 and ***p < 0.001 compared to control (Dunnett's test). Data shown is representative of four independent experiments. B) Lung CD11c+ dendritic cells were extracted from mice exposed to cigarette smoke for ≥4 weeks in a smoking chamber and adjusted to a concentration of 1 × 106/ml in media containing 5 ng/ml of murine GM-CSF. Viability of immature and LPS-matured lung dendritic cells from 4 cigarette smoke exposed mice and 4 control mice was performed using XTT assay following 24 hours of incubation. iDC = immature DC. 2-way ANOVA p < 0.01. C) Spleen CD11c+ dendritic cells were also extracted from mice exposed to cigarette smoke for ≥4 weeks in a smoking chamber and adjusted to a concentration of 1 × 106/ml in media containing 5 ng/ml of murine GM-CSF. Viability of immature and LPS-matured spleen dendritic cells was also performed using XTT assay following 24 hours of incubation. Data shown in B and C are representative of 2 independent experiments. D and E) Human dendritic cells (1 × 106/ml in complete media) were incubated with 0-1000 ng/ml nicotine for 24 or 48 hours prior to determination of viability with an XTT assay. Data shown in D and E are representative of 3 independent experiments. Data shown are means ± SEM. *p < 0.05, **p < 0.01.