CD163 expression is induced by <i>Leishmania</i> infection of monocyte/macrophages.

<p>(A) Gating strategy for analysis of monocyte/macrophage phenotypes. PBMC were collected from healthy donors and the adherent cells cultured for 5 days in RPMI 1640 plus 20% FBS. The cells were infected with <i>L</i>. <i>amazonensis-</i>GFP (10 parasites: 1 macrophage), two isolates of <i>L</i>. <i>infantum</i> CellTracker stained (5:1) or BCG CellTracker stained (2:1), and incubated with antibodies specific for macrophage surface molecules prior to analysis by flow cytometry. (B) Surface CD163 expression was quantified after 24h in infected, GFP+ cells (in duplicate, n = 5 experiments). (C) The macrophages were analyzed by FlowJo software (in duplicate, n = 6 experiments). Green and black bars represent, respectively, the infected, GFP+ and uninfected, GFP- cells after 24h of exposure to <i>L</i>. <i>amazonensis</i> GFP-expressing strain. White bars are the non-exposed group (unstimulated). The mean ± SD parental percentage of the cell populations (B and C) were compared by Friedman paired test with Dunn’s post test. (C) MFI analysis of GFP in the different populations according to the surface phenotype, 24h after infection (n = 7 experiments in duplicate). The mean ± SD MFI of each population was compared by Friedman paired test with Dunn’s post test. (D) Spearman correlation analysis between CD163-PE and <i>Leishmania-</i>GFP fluorescence for each cell of the analysis (one representative of 7 experiments in duplicate).