Vγ9Vδ2-T–cell recognition and lysis of CLL cells. (A) Representative plot of geometric mean fluorescence intensity (gMFI) of CD25 expression measured after a 36-hour coculture of HC Vγ9Vδ2-T cells with CLL cells (solid red line), allogeneic healthy B cells (dashed blue line), or Vγ9Vδ2-T cells alone (light green shading). (B) As in (A), scatter plot summarizing results for 6 donors. (C-E) Cell death of carboxyfluorescein succinimidyl ester–labeled target cells after overnight coculture with Vγ9Vδ2-T cells, measured by MitoTracker Orange and TO-PRO-3. (C) Specific lysis of CLL cells after coculture with Vγ9Vδ2-T cells from HCs at the indicated effector-to-target ratios (n = 9). Specific lysis was calculated as (% cell death in stimulated cells) − % cell death in unstimulated cells)/(% viable cells in unstimulated cells) * 100. (D) Healthy Vγ9Vδ2-T cells were treated for 2 hours with 100 nM concanamycin A (CMA) or dimethyl sulfoxide and washed before coculture with CLL cells at a 1:1 ratio (n = 9). (E) Specific lysis of allogeneic CLL and Daudi cells after coculture with Vγ9Vδ2-T cells from CLL patients (n = 6) or HCs (n = 9) at a 1:1 ratio. (F) CD69 expression on Vγ9Vδ2-T cells from CLL patients (n = 8) or HCs (n = 4) after overnight coculture with Daudi cells at a 1:5 ratio. Data are mean and SEM. *P < .05, **P < .01, ***P < .001, ****P < .0001, 1-way ANOVA, followed by the Dunnett (B) or Bonferroni (E-F) correction; paired t test (D).