Gene expression profiling of plasmacytoid dendritic cells from cutaneous lymph nodes

Plasmacytoid dendritic cells wre isolated from cutaneous lymph nodes of control C57BL/6 mice and used for microarray analysis. Plasmacytoid dendritic cells were isolated from cutaneous lymph nodes (CLNs) as previously described (Henri et al. 2010). Briefly, CLNs were cut into small pieces and digested for 20 min at room temperature with a mixture of type II collagenase (Worthington Biochemical) and DNase I (Sigma-Aldrich). The resulting cell suspension was treated with 5 mM EDTA to disrupt DC-T cell conjugates. After eliminating undigested material, light-density cells were enriched by centrifugation on an Optiprep solution (d = 1.32 g/mL, Abcys).