Cytokine production by cultured DCs from healthy controls, disease controls, and the patient. (A) The cell surface expression of monocyte-specific (CD14) and DC-specific (DC-SIGN) surface molecules was analyzed on cells immediately after selection on CD14 beads (post-CD14⁺ purification) and after a 7–8-d culture in GM-CSF + IL-4 with the addition of TNF for the final 24 h of culture (post-DC differentiation). The isotype control is shown in gray, and the specific staining for the healthy control, disease control, and patient cells are shown in black, blue, and red, respectively. (B) Representative qRT-PCR analysis of inflammatory messenger RNA (mRNA) expression by stimulated DCs. DCs isolated from control (black), disease controls (blue), or the patient (red) were stimulated with 100 ng/ml LPS (left) or 100 µg/ml curdlan (right), and messenger RNAs were analyzed by qRT-PCR with normalization to GAPDH. All the PCRs were performed twice with independent cDNA preps. The disease controls had mutations in BTK. Note the difference in scale in the response to LPS versus curdlan.