Figure 4.

Subset-specific assessment of intracellular TNF, IL1b, and IL6 production in circulating blood monocytes after incubation in whole blood for six hours with PBS (control), mCRP, and LPS. Brefeldin A was used to inhibit the protein transport from the endoplasmic reticulum to the Golgi complex. Both monocyte subsets showed a rapid increase of TNF-positive monocytes in a subset-specific manner upon LPS and mCRP stimulation with non-classicals monocytes being the main source of TNF (A, B). For IL1b, a subset-specific upregulation of intracellular production, predominantly in classical monocytes, was found both following mCRP and LPS stimulation. Within classical monocytes, IL1b production was significantly higher upon mCRP stimulation (C, D). In contrast, the intracellular upregulation of IL6 production markedly increased upon both reagents and showed a subset-specific pattern only following LPS incubation (E, F). ns, not significant; ***p < 0.001; ****p < 0.0001.