Fig. 3

SPADE (Spanning-tree Progression Analysis of Density-normalized Events) analysis. T-cell phenotype was evaluated by using a SPADE algorithm based on the expression of different markers measured by flow cytometry on whole blood samples from patients with septic shock at day 3 after the onset of shock (D3, n = 17) and in healthy volunteers (HVs) (n = 14). CD4⁺ and CD8⁺ T cells from patients and donors were clustered in nodes on the basis of their similarities of expressions of CD127, CD38, HLA-DR, PD-1, FoxP3, and CD25. One tree containing four nodes was built for CD4⁺ T lymphocytes and one tree containing three nodes for CD8⁺ T cells. The medians of fluorescence intensity (MFIs) of the different markers for each node are represented on CD4⁺ and CD8⁺ T cells in patients and in HVs. Data are presented as Tukey boxplots. Node 4 of the CD4⁺ T cells, with a CD127^lowCD25^highFoxP3^high phenotype, corresponds to regulatory T cells. Node 3 corresponding to CD127^lowPD-1^high cells has been highlighted in both CD4⁺ and CD8⁺ T cells