Nucleotides Regulate Secretion of the Inflammatory Chemokine CCL2 from Human Macrophages and Monocytes
Figure 5
Primary human peripheral blood monocytes express P2Y2 but display variable CCL2 secretion to UTP. (a) Human PBMCs were labelled with conjugated CD14-FITC and rabbit anti-P2Y2 followed by goat anti-rabbit Alexa 647 using an intracellular staining protocol. (b) Human CD14 positive magnetically isolated monocytes were plated on poly-D-lysine plates and loaded with Fluo-4AM. UTP induced a transient calcium response at both 10 μM (orange) and 100 μM (blue). Calcium measurements were recorded using a Fluostar Optima plate reader at 37°C. (c) Human CD14+ monocytes were plated at 5 × 105 cells/well and stimulated with media, LPS 100 ng/mL, or 10 μM UTP for 4 hours. Each symbol represents a different donor ( donors in total). (d) Human CD14+ monocytes stimulated for 6 hours with media, LPS 100 ng/mL, 10 μM UTP, or 100 μM UTP. Each symbol represents a different donor ( different donors).
