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Figure 1 | Journal of Hematology & Oncology

Figure 1

From: TIM-3/Gal-9 interaction induces IFNγ-dependent IDO1 expression in acute myeloid leukemia blast cells

Figure 1

NK/AML co-culture induces TIM-3/Gal-9-dependent IDO-1 activation. BM samples from children with AML at diagnosis were processed by Ficoll-Paque Plus to obtain BM mononuclear cells. PBMC cells processed from buffy coat preparations of healthy donors were cultured for 10 days on a feeder layer of irradiated RPMI 8866 cells to obtain polyclonal expansion of NK cells. (A) IDO1 and IRF1 expression on protein extracts from 24-h culture (5:1 ratio) of AML blasts and AML blasts pre-treated for 1 h with anti-Gal-9 antibody with NK cells compared with NK cells alone. GAPDH Ab was used as loading control. Supernatants were collected and used for the measurement of kynurenine levels by RP-HPLC in the different culture conditions. Results shown are representative of 3 AML samples. (B) ELISA assay for IFNγ release was performed in the supernatants of experimental co-cultures. Bars are representative of mean values and standard deviation recorded in three independent experiments performed in duplicate (*P < 0. 05).

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