Transcriptional effects of soluble CD40 ligand on human naïve B cells

We performed microarray analysis to derive gene signatures down-stream of soluble CD40 ligand stimulation in human naive B cells. Naïve B cells were purified from healthy donor PBMC using negative selection beads (Miltenyi) and cultured with sCD40L at 2.5ug/ml for 6hr before microarray analysis. In the same study, cells were also harvested at day 5 post-stimulation to confirm sCD40L-induced B cell activation and proliferation. FACS analysis confirmed soluble CD40L induced up-regulation of CD86 and CD69 at 24hr. B cell proliferation was measured at day 4 post-stimulation by EdU incorporation. Naïve B cells were purified from peripheral blood of 6 healthy donors. Naïve B cells were then stimulated with sCD40L or mock-treated, and harvested at 6hr for microarray analysis. 2-4 replicates were available per donor and condition.