Cd163

CD14, CD68, CD11b, CD11c, CD83 and CD1a fluorochrome-conjugated antibodies (BD Biosciences) were used to stain the immature macrophages in the dark at 4 °C for 15 min. After co-culture experiments for 72 h, immature macrophages were obtained by incubating with CD163 (Miltenyi Biotec), CD204 (Miltenyi Biotec), CD206 (Miltenyi Biotec) and B7-H1 (eBioscience) fluorochrome-conjugated antibodies in the dark at 4 °C for 15 min, followed by incubation with iNOS antibody (Abcam) and fluorescently-labeled secondary antibody. Cells were analyzed on a FACScan flow cytometer (BD Biosciences), and data were interpreted using the Flowjo software (Tree Star). Each experiment was performed in triplicate.

Peripheral blood mononuclear cells collected at admission were stained for CD3, DC-SIGN (Beckman Coulter, Brea, CA, USA) CD8, CD40, HLA-DR (BioLegend, San Diego, CA, USA), CD14, CD16, CD19, CD56, CD86, CD209 (BD Biosciences), CD163 (Miltenyi, Bergisch Gladbach, Germany), and Live/Dead Fixable Yellow (Thermofisher, Waltham, MA, USA) (details in S1 Table). Data were acquired on a Fortessa X20 flow cytometer (BD Biosciences) and analyzed were using Diva software (BD Biosciences). The Excyted pipeline was used to normalize and cluster the data (https://github.com/maximemeylan/Excyted). Intensity values of events gated from live cells were normalized using the Logicle transformation. Unsupervised clustering and Uniform Manifold Approximation and Projection (UMAP) were computed with 10.000 events for each sample using k = 30 [27 ].