Intracellular expression of cytokines in BM DCs terminally differentiated by LPS or CD40 ligation.

(A) Detection of intracellular cytokines by flow cytometry in CD11c⁺ immunobead-sorted CD86⁺ BM DCs (control) or terminally differentiated with LPS or anti-CD40 IgM mAb. DCs treated with irrelevant IgM exhibited a pattern similar to that of the control group (not shown). Expression of IFN-α was also investigated in CD86⁺ BM DCs infected with a rAd encoding no transgene (MOI = 100) used as positive control. DC were double labeled with FITC anti-CD11c and specific PE anticytokine mAbs. Figures inside quadrants indicate percentages of cells. (B) Detection of TGF-β1 by ELISA in 24-hour culture supernatants of FACS-sorted CD86⁺mature DC (2 × 10⁶ cells/well) incubated with LPS, or anti-CD40 IgM mAb, or irrelevant IgM (control). Results are representative of 3 independent experiments.