Transcriptional effects of soluble CD40 ligand on human immature dendritic cells

We performed microarray analysis of sCD40L-stimulated iDC to derive a signature of CD40 activation. Human monocytes from normal healthy donors were differentiated to iDCs with GM-CSF and IL4. FACS analysis demonstrated the immature status of these cells, illustrated by low expression of CD80, CD40, and CD86. We confirmed that sCD40L induces the maturation of DCs, characterized by higher expression of CD80, HLA-DR, CD86, CD83 and CD40 and secretion of pro-inflammatory cytokines at 24hr post-stimulation. Cells were harvested at 1, 3 and 24hr post-stimulation for microarray analysis. Immature dendritic cells were derived from 5 healthy donors. Cells were treated with vehicle control or soluble CD40L for 1, 3, or 24hrs before RNA extraction and microarray processing with Affymetrix HT-HGU133 2 Plus PM arrays.