Gene expression in β-glucan-stimulated human DCs.

<p>(A) Human monocyte-derived DCs were cultured for 24 h with or without particulate β-glucan or LPS. Protein secretion was measured by ELISA in culture supernatants. Results are mean ± SEM, n = 12 to 31. Statistics: Wilcoxon signed-rank test (TNF, IL-6, IL-12p40, and IL-10) or Tobit model analysis (IL-1β, IL-23, IL-1α, IL-18, and IL-12p70). (B and C) Human monocyte-derived DCs were cultured in the presence (solid lines) or absence (dashed lines) of particulate β-glucan for the times shown. Transcript accumulation was determined by absolute Real-Time qRT-PCR using total RNA and primers detecting mature transcripts (Table S2 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114516#pone.0114516.s001" target="_blank">File S1</a>). Protein secretion was measured by ELISA in the culture supernatants. Results are mean, n = 6 to 8 for transcripts, n = 5 to 13 for proteins. (D) Gene expression in human monocyte-derived DCs was evaluated by microarrays after activation with β-glucan for the indicated times. The heat map shows differentially expressed [≥2-fold change in expression (FDR <0.1) in at least one time point compared to the pre-stimulation level] genes (rows) in cells from three different donors (columns). The mRNA profiles were hierarchically clustered and subdivided into six groups (leftmost column of each heat map) according to the time when β-glucan controlled their expression. (E) Cluster models (Cluster ONE Cytoscape plugin) illustrating the pathways associated with the 3 groups of genes induced by β-glucan (GO pathway analysis) (left panels) and their predicted TFs (TRANSFAC database) (right panels). Each node of the pathway clusters depicts a single pathway. Similar pathways were grouped under a single representative name. Each node of the TF clusters indicates a redundant name or a predicted binding site for the listed TFs. The size of the nodes indicates the significance of the association of the pathway or the potential binding site for a TF.