Figure 3.

EC0746 has an immunomodulatory effect on folate-receptor-expressing rat TG-macs. Rat TG-macs were treated with media only, 100 nM EC0746 ± 10 μM folic acid (FA), or FA alone (10 μM) for 2 hours followed by a 70-hour chase. In comparison, the cells were also treated with 100 nM free aminopterin (AMT) and methotrexate (MTX) for 72 hours. At 24 hours before the end of incubation, all cells were stimulated with lipopolysaccharide (LPS) (5 μg/ml) plus IFNγ (100 ng/ml). The cytokines/chemokines produced in culture supernatants were detected using a rat cytokine array. (a) Cytokine release profiles of rat TG-macs stimulated with LPS and IFNγ with or without drug treatment. (b) Cytokine array map. (c) Mean pixel intensity (y axis) determined for each array position and plotted for the 11 products, which were detected at three times above background levels and in at least three independent experiments. Data shown are mean ± standard error of the mean. *P < 0.05 when compared with its corresponding cytokine level in the media only sample. CINC, cytokine-induced neutrophil chemoattractant; LIX, LPS-induced CXC chemokine; MIG, monokine induced by IFNγ; MIP-1α, macrophage inflammatory protein-1α; RANTES, regulated upon activation, normal T-cell expressed and secreted; sICAM, soluble intracellular adhesion molecule; TIMP-1, tissue inhibitor of metalloproteinase 1; VEGF, vascular endothelial growth factor.