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Figure 2 | Journal of Translational Medicine

Figure 2

From: Indoleamine 2,3-dioxygenase 1 (IDO1) activity correlates with immune system abnormalities in multiple myeloma

Figure 2

IDO1 activity in MM BMSC. Panel A: BMSC were generated as previously detailed [33] and were either left untreated or activated with 100 IU/ml IFN-γ for 24 hours. BMSC were fixed, permeabilized and stained with anti-IDO and anti-CD166 mAb. One representative experiment out of 5 with similar results is shown. Panel B: BMSC from 5 representative patients with MM and skin fibroblasts from 6 healthy subjects were either left untreated (black dots) or activated with 100 IU/ml IFN-γ for 24 hours (red dots) before measuring KYN and tryptophan level in culture supernatants. Comparisons between untreated and IFN-γ-stimulated samples were performed with the Wilcoxon matched-paired signed rank test. *p<0.01 compared with untreated samples. Panel C: Supernatants of BMSC cultures were transferred (20% v/v) to an MLR containing allogeneic naïve CD4+ T cells and third-party monocytes (at a fixed 1:3 ratio). IL-2 was provided to the cultures as a Treg growth factor. After 6 days, cells were harvested and labeled with anti-FoxP3 and anti-CD25 mAb to monitor the acquisition of a regulatory phenotype. A representative experiment out of 3 with similar results is shown. Markers were set according to the proper isotypic control (not shown). Rectangular gates were set to include CD4+CD25hiFoxP3hi T cells.

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