Altered metabolic response in IFNAR^−/− DCs.

<p> (A) Bone marrow mixed-chimera mice were injected with 50 µg poly IC IP. 14 h later, DCs were FACS sorted based on CD45.2 expression and analyzed for gene expression with Illumina chips. Heat map represents expression values (2-fold change and FDR<0.05) of poly IC-treated WT (green), IFNAR^−/− (orange) versus untreated DCs (blue). Each column represents an individual sample. (B) Pathway analysis was performed using GSEA (FDR<0.25). Table represents the top ten pathways upregulated in IFNAR^−/− versus WT DCs.</p

Changes in mitochondrial function after poly IC stimulation.

<p>WT/IFNAR bone marrow mixed-chimera mice were injected with 50 µg poly IC IP. 14 h later, spleen were harvested and incubated with the membrane potential-dependent stain MitoTracker Red CMXRos (A) or MitoTracker Green FM (B) for 20 min at 37°C. Cell staining was evaluated in CD11c⁺MHCII⁺B220⁻DX5⁻ DCs by flow cytometry. Histogram is representative of three independent experiments. (C) As (A) and (B) but after 14 h, CD11c⁺MHCII⁺B220⁻DX5⁻ DCs were sorted and seeded in a Seahorse XF-24 analyzer. OCR was determined in real time after the addition oligomycin, FCCP, and antimycin-A/rotenone. (D) ECAR values. Data represent means ± SD of quadruplicates. Data are representative of three experiments. (E) WT mice were injected with 50 µg poly IC IP 18 h later intracellular expressions of iNOS was evaluated by FACS. As positive control, monocyte-derived DCs were stimulated with LPS. Histogram are representative of three independent experiments. (F) As in (A), but WT were injected with 50 µg poly IC IP, spleen was harvested 4 h later and further incubated in the presence of SEITU ON. Histogram is representative of three independent experiments. (G) Chimera mice were stimulated with poly IC 4 h in vivo. Total RNA was isolated from sort-purified CD11c⁺MHCII⁺ DCs. Relative expression of HIF1α was analyzed by real time (RT)-PCR or by intracellular staining. mRNA levels in resting DCs were set to 1.</p

Immune systems analysis in the hyperacute window.

<p>(A) Gene expression profiles of the 16 genes with the greatest difference in expression between the hyperacute window and 24 hours postinjury in the Critical versus Control transcriptome analysis. (B) Immune module analysis of the critical response to injury. Again, there was a differential response between the hyperacute response and the later time points. There was overexpression in modules associated with inflammation and of neutrophil and natural killer (NK) cell modules. There were mixed over- and underexpression in modules related to T cells and B/plasma cells and underexpression of monocyte-related modules (contrary to the deconvolution data in <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1002352#pmed.1002352.g002" target="_blank">Fig 2B</a>).</p

Patient demographics.

<p>Patient demographics.</p