Extended Data Fig. 3: Immunosuppression in the late immunosuppressive phase following LPS administration.

a, Heatmap of changes in flu-like symptoms, body temperature, and heart rate after the first and second intravenous challenge with 2 ng kg⁻¹ LPS in healthy volunteers (n = 7). b, Heatmap of differences in log2(fold-change) blood concentrations of cytokines TNF, IL-10, CCL4, CCL3, IL-6, CXCL8, CCL2, IL-1RN between the first and second intravenous challenge with 2 ng kg⁻¹ LPS in healthy volunteers (n = 7). c, Heatmap of changes in blood leukocyte (subtypes) after the first and second intravenous challenge with 2 ng kg⁻¹ LPS in healthy volunteers (n = 7). d, Scatterplot of mean normalized gene expression profiles of blood CD14⁺ monocytes from LPS-challenged volunteers (n = 3) obtained at 4 h, 8 h, 24 h, and d7 that were ex vivo stimulated with LPS (10 ng mL⁻¹) vs. CD14⁺ LPS-stimulated monocytes obtained at BL. Orange dots indicate semi-suppressed genes (-1>log2[fold-change]>-2) and red dots indicate suppressed genes (-2>log2[fold-change]). e, Heatmap of average log2(fold-change) difference in cytokine (TNF, IL6, IL1β, CCL4, IL10, IL1RN) production by blood CD14⁺CD16⁻ classical monocytes (cMonos) obtained at 4 h and d7 after LPS administration in healthy volunteers (n = 7) that were ex vivo stimulated with Pam3Cys, Poly:IC, LPS, Flagelline, R848, E. coli, S. aureus, P. aeruginosa, C. albicans, and A. fumigatus compared with cytokine production of cMonos obtained from the same subjects at baseline (before LPS administration) stimulated with the same ligands. f, violin plot of log2(fold-change) difference in gene expression of blood CD14⁺ monocytes from LPS-challenged volunteers (n = 3) obtained at 4 h, 8 h, 24 h, and d7 that were ex vivo stimulated with LPS (10 ng mL⁻¹) vs. CD14⁺ LPS-stimulated monocytes obtained at BL. Genes are clustered based on their dynamics over time (see main text and Fig. 1l). g, Scatterplot of log2(normalized expression) of IFN-I signaling pathway genes in blood CD14⁺ monocytes from LPS-challenged volunteers obtained at d7 that were ex vivo stimulated with LPS (10 ng mL⁻¹, y-axis, mean expression of n = 3) compared to LPS-stimulated CD14+ monocytes form the same volunteers obtained at baseline (x-axis, mean expression of n = 3). h, Schematic representation of ex vivo BST2 flow cytometry experiments in immunosuppressed pan monocytes (containing all subsets) i, Representative example of flow cytometric analysis of monocyte subsets immediately after isolation, and after 24 h of incubation with or without LPS (10 ng mL⁻¹). j, Overlay of BST2 mean fluorescence intensity (MFI) in cMonos obtained at baseline and at d7 after 24 h of ex vivo stimulation with LPS (10 ng mL⁻¹). k, Bar plot of BST2 MFI in cMonos (n = 3) obtained at baseline and at d7 after 24 h of ex vivo stimulation with LPS (10 ng mL⁻¹). Bar plot in k presented as mean values +/− SEM. The P value calculated using a paired two-sided t-test.