SINEs do not alter T cell or NK cell viability but negatively influences IL-6 and IL-10 production. (A) CD3⁺ T cells (N = 6) from normal volunteers were incubated with or without 1μM of KPT-185 for 48 hours. Cells were stimulated using an anti-CD3 T-cell activation plate for additional 24 hours. Cells viability (ann/PI negative cells) was measured by annexin-V/PI flow cytometry and was calculated relative to time-matched untreated controls. (B) CD56⁺ NK cells (N = 6) from normal volunteers were incubated with or without KPT-185 for 72 hours. Viability was measured by annexin-V/PI flow cytometry and was calculated relative to time-matched untreated controls. (C-E) Supernatant from anti-CD3 stimulated T cells treated with or without 1μM of KPT-185 for 48 hours was collected and IL-6, IL-10, and TNF-α production were measured by ELISA. (F) ADCC against CLL cells was measured using KPT-185 or KPT-251–treated NK cells (12 hours) from normal volunteers and CLL cells at 6.25:1, 12.5:1, and 25:1 effector to target ratio (E:T) in the presence or absence of 10 μg/mL ofatumumab, alemtuzumab, or trastuzumab. Columns are averages of triplicate wells, and are representative of 3 independent experiments; bars represent SD. (G) NK directed cytotoxicity against K562 cells was measured using KPT-185 or KPT-251–treated NK cells (12 hours) from normal volunteers and K562 cells at E:T ratios of 6.25:1, 12.5:1, and 25:1. Columns are averages of triplicate wells and are representative of 3 independent experiments; bars represent SD.