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Figure 4 | Journal of Translational Medicine

Figure 4

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

Figure 4

IDO+myeloma cells induce a Tr1/Th2 profile and suppress Th1 cytokines in allogeneic CD4+T cells. Panels A and B: Highly pure CD138+ PC from patients with MM were used for MLTC, as detailed in Materials and Methods. The allogeneic CD4+CD25- T cells were pre-loaded with CFSE to monitor cell division. Exogenous IL-2 was provided to the MLTC at 100 IU/ml (final concentration). In selected wells, D,L- 1MT was added at 200 μM (final concentration) to inhibit IDO activity. After 6 days in culture, cells were harvested and labeled with anti-FoxP3 and anti-CD25 mAb. Quadrant markers were set according to the proper isotypic control. A representative experiment is shown in panel A, whereas results are summarized in panel B. Panel C: CFSE dilution in responder CD4+CD25- T cells was expressed in terms of mean fluorescence intensity of the dividing cell population (MFI). The bars depict the median and interquartile range recorded in 3 independent experiments performed in duplicate. The black column denotes T-cell cultures that were stimulated with anti-CD2+anti-CD3+anti-CD28 mAb in the absence of MM-induced Treg cells. Panel D: MM cells from KYNhi patients were seeded in a MTLC as above detailed. After 6 days in culture, T cells were collected and used for intracellular cytokine staining. The percentage of IL-2, and IFN-γ-expressing T cells shown in panel E is representative of 3 independent experiments. Quadrant markers were set according to the proper isotypic control.

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