Anti-CD40 activated TC-derived BMDCs secrete IFN-γ.

<p><b>A</b>. IFN-γ gene expression in BMDCs stimulated or not with anti-CD40 Ab for 24 h. Message expression was normalized to <i>Gapdh</i> and expressed relative to the mean values for unstimulated B6 BMDCs for each gene. (N = 7 per group). <b>B</b>. Intracellular IFN-γ expression expressed as mean fluorescence intensity (MFI) in BMDCs stimulated or not with anti-CD40 Ab for 24 h (N = 4 per strain). <b>C</b>. Representative FACS plots showing intracellular IFN-γ staining in CD11c^hi viability dye-negative anti-CD40 stimulated BMDCs (corresponding to the graph in B). The middle and right-side overlays shown BMDCs stimulated with IL-12p70 and IL-18 as controls. The solid black lines correspond to TC DCs and the broken lines correspond to B6 DCs. The filled gray histograms show the isotype control. <b>D</b>. B6 B cell proliferation measured as CSFE dilution in response to anti-CD40-activated TC BMDC (red) or B6 BMDC (blue) supernatant. The profile of B cell alone is shown as control. Representative FACS overlays and quantitation. <b>E</b>. Differential expression of IFN-γ-inducible genes in the B cells shown in <b>D</b>. The graph shows the TC over B6 fold expression for all the IFN-γ-inducible genes that are expressed as a p<0.01 different level between the B cells activated with the two different types on activated BMDCs.