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Figure 1: Tim-3 and PD-1 blockade could not improve the degranulation of NK cells in CLL patients. Magnetic-activated cell sorting-isolated NK cells from CLL patients (n = 18) were treated with human anti-Tim3 and/or anti-PD-1 (alone or combined) and isotype control antibodies. Cells were stimulated with IL-2 for 24 h and then treated with PMA/ionomycin cocktail and finally co-cultured with K562 cell line as target (effector: target ratio 1:1). Overnight incubation was performed with fluorochrome-conjugated anti-CD107a and its irrelevant isotype control antibodies. In flow cytometry analysis, NK cells were initially gated, and then, the expression of CD107a was determined. No significant differences were observed among control and blocked groups (P > 0.05). (a) A representative flow cytometric dot plot gating on population of NK cells and the flow cytometry histograms for CD107a expression obtained from a CLL patient are represented. (b) CD107 expression on NK cells from all CLL patients. Graph represents the mean ± SD. Tim-3, CLL = Chronic lymphocytic leukemia, SD = Standard deviation, IL-2 = Interleukin-2, NK = Natural killer cell

Figure 1: Tim-3 and PD-1 blockade could not improve the degranulation of NK cells in CLL patients. Magnetic-activated cell sorting-isolated NK cells from CLL patients (<i>n</i> = 18) were treated with human anti-Tim3 and/or anti-PD-1 (alone or combined) and isotype control antibodies. Cells were stimulated with IL-2 for 24 h and then treated with PMA/ionomycin cocktail and finally co-cultured with K562 cell line as target (effector: target ratio 1:1). Overnight incubation was performed with fluorochrome-conjugated anti-CD107a and its irrelevant isotype control antibodies. In flow cytometry analysis, NK cells were initially gated, and then, the expression of CD107a was determined. No significant differences were observed among control and blocked groups (<i>P</i> > 0.05). (a) A representative flow cytometric dot plot gating on population of NK cells and the flow cytometry histograms for CD107a expression obtained from a CLL patient are represented. (b) CD107 expression on NK cells from all CLL patients. Graph represents the mean ± SD. Tim-3, CLL = Chronic lymphocytic leukemia, SD = Standard deviation, IL-2 = Interleukin-2, NK = Natural killer cell