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Figure 6 | Journal of Translational Medicine

Figure 6

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

Figure 6

HGF modulates IDO in MM cell lines. Panel A: Mo-DC were differentiated from circulating monocytes of healthy donors with GM-CSF+IL-4 (7 days; immature Mo-DC or iDC) and then matured with TNF-α (2 additional days; mature DC or mDC), as already published [26]. Mature Mo-DC were routinely CD14dim/negCD1a+, as determined by FACS analysis. Panel B: Mo-DC differentiated in the presence of exogenous HGF at 20 ng/ml and then matured with TNF-α up-regulated IDO protein. Panel C: U266 and MOLP-8 MM cells were stimulated with either IFN-γ or HGF for 24h. Messenger RNA levels for IDO were measured with quantitative RT-PCR relative to β-actin. Bars depict mean and SD recorded in 3 independent experiments. Panel D: U266 and MOLP-8 MM cells were stimulated with 20 ng/ml HGF for 24 hours. Cell lysates were immunoblotted with phosphorylation-specific antibodies against AKT (Ser473) [62]. SU11274, a selective MET inhibitor, was provided at 100 nM to the cultures. IFN-γ at 100 IU/ml was used as a prototypical inducer of IDO. Panel E: Tryptophan and KYN concentrations were measured in supernatants of U266 and MOLP-8 MM cells that were stimulated with 20 ng/ml HGF either in the presence or absence of 1MT. Results are representative of 3 independent experiments run in duplicate.

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