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. 2014 Nov 20;8(11):e3330. doi: 10.1371/journal.pntd.0003330

Figure 1. IDO Inhibition increases the fungal loads of macrophages and inhibits kynurenines and IDO mRNA production.

Figure 1

(A) IFN-γ-primed and unprimed peritoneal macrophages from A/J and B10.A mice strains were treated with 1MT (1 mM) or left untreated and subsequently infected with P. brasiliensis yeasts in a macrophage∶yeast ratio of 25∶1 for 2 h. Infected and uninfected macrophages were then cultivated for 48 h at 37°C in 5% CO2. The monolayers were lysed with distilled water, and 100 µl of cell homogenates were assayed for the presence of viable yeasts by a CFU assay. (B, C) Supernatants from macrophage cultures were used to determine the levels of nitrite and kynurenines. (D) In the same experimental conditions, peritoneal macrophages of A/J and B10.A mice were mixed with TRizol reagent for RNA extraction and then submitted to qRT-PCR assays for mRNA IDO evaluation (D). Data are the means ± SEM of quintuplicate samples from one experiment representative of 3 independent determinations (*p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001).