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Figure 3: Tim-3 and PD-1 blockade could not alter the production of IFN-γ and TNF-α by NK cells in CLL patients. Magnetic-activated cell sorting-isolated NK cells from CLL patients (n = 18) were treated with human anti-Tim-3, anti-PD-1 (alone or combined), and isotype-matched control antibodies. Cells were stimulated with IL-2 for 24 h and then treated with PMA/ionomycin cocktail and finally co-cultured with K562 target cells (effector:target ratio 1:1). Following overnight incubation, culture supernatants were collected to measure the levels of TNF-α (a) and IFN-γ (b) by ELISA. The amount of secreted cytokines among control and blocked groups was not significantly different (P > 0.05). Both graphs represent the mean ± SD. IFN-γ = Interferon gamma, TNF-α = Tumor necrosis factor-alpha, CLL = Chronic lymphocytic leukemia, IL-2 = Interleukin-2, SD = Standard deviation, NK = Natural killer cell

Figure 3: Tim-3 and PD-1 blockade could not alter the production of IFN-γ and TNF-α by NK cells in CLL patients. Magnetic-activated cell sorting-isolated NK cells from CLL patients (<i>n</i> = 18) were treated with human anti-Tim-3, anti-PD-1 (alone or combined), and isotype-matched control antibodies. Cells were stimulated with IL-2 for 24 h and then treated with PMA/ionomycin cocktail and finally co-cultured with K562 target cells (effector:target ratio 1:1). Following overnight incubation, culture supernatants were collected to measure the levels of TNF-α (a) and IFN-γ (b) by ELISA. The amount of secreted cytokines among control and blocked groups was not significantly different (<i>P</i> > 0.05). Both graphs represent the mean ± SD. IFN-γ = Interferon gamma, TNF-α = Tumor necrosis factor-alpha, CLL = Chronic lymphocytic leukemia, IL-2 = Interleukin-2, SD = Standard deviation, NK = Natural killer cell