Pharmacological inhibition of eicosanoids and platelet-activating factor signaling impairs zymosan-induced release of IL-23 by dendritic cells

Abstract

The engagement of the receptors for fungal patterns induces the expression of cytokines, the release of arachidonic acid, and the production of PGE2 in human dendritic cells (DC), but few data are available about other lipid mediators that may modulate DC function. The combined antagonism of leukotriene (LT) B4, cysteinyl-LT, and platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) inhibited IL23A mRNA expression in response to the fungal surrogate zymosan and to a lower extent TNFA (tumor necrosis factor-α) and CSF2 (granulocyte macrophage colony-stimulating factor) mRNA. The combination of lipid mediators and the lipid extract of zymosan-conditioned medium increased the induction of IL23A by LPS (bacterial lipopolysaccharide), thus suggesting that unlike LPS, zymosan elicits the production of mediators at a concentration enough for optimal response. Zymosan induced the release of LTB4, LTE4, 12-hydroxyeicosatetraenoic acid (12-HETE), and PAF C16:0. DC showed a high expression and detectable Ser663 phosphorylation of 5-lipoxygenase in response to zymosan, and a high expression and activity of LPCAT1/2 (lysophosphatidylcholine acyltransferase 1 and 2), the enzymes that incorporate acetate from acetyl-CoA into choline-containing lysophospholipids to produce PAF. Pharmacological modulation of the arachidonic acid cascade and the PAF receptor inhibited the binding of P-71Thr-ATF2 (activating transcription factor 2) to the IL23A promoter, thus mirroring their effects on the expression of IL23A mRNA and IL-23 protein. These results indicate that LTB4, cysteinyl-LT, and PAF, acting through their cognate G protein-coupled receptors, contribute to the phosphorylation of ATF2 and play a central role in IL23A promoter trans-activation and the cytokine signature induced by fungal patterns.

Keywords: 12-HETE (PubChem CID: 5353272); 12-HETE-d8 (PubChem CID: 1436); Acetyl-CoA (PubChem CID: 444493); Arachidonoyl-CoA (PubChem CID: 52922027); Cytokines; Darapladib (PubChem CID: 9939609); Dendritic cells; EDC (2-TEDC, PubChem CID: 6069552); Fungus; Leukotriene B(4) (PubChem CID: 5280492); Leukotriene B(4)-d4 (PubChem CID: 53394387); Leukotriene C(4) (PubChem CID: 5280493); Leukotriene E(4) (PubChem CID: 5280879); Lipid mediators; Lyso-PAF C16:0 (PubChem CID: 3983); Montelukast (PubChem CID: 5281040); PAF C16:0-d4 (PubChem CID: 53394395); PAF C18:0 (PubChem CID: 2733535); PGE(2) (PubChem CID: 5280360); Platelet-activating factor (PubChem CID: 108156); Signal transduction; U75302 (PubChem CID: 6449854); WEB2086 (Apafant, PubChem CID: 65889).