Ibrutinib promotes TH1 phenotype in Vγ9Vδ2-T cells. (A-B) CD19-depleted PBMCs were treated with 0, 10, or 100 nM ibrutinib for 30 minutes and subsequently cocultured with ABP-pretreated (25 μM pamidronate for 2 hours) Daudi cells. During the last 6 hours of coculture, brefeldin A and GolgiStop were added to measure cytokine production (A) and CD107a expression (B) in Vγ9Vδ2-T cells (n = 10). (C) Cytokine production as in (A) after pretreatment with 0, 10, or 100 nM CC-292 (n = 6). Data are mean and SEM. (D) Pull-down with biotinylated ibrutinib coupled to avidin agarose; uncoupled avidin agarose was used as a control. Lysates from healthy Vγ9Vδ2-T cells or control Mec-1 B cells were treated with 1 μM ibrutinib or 1 μM CC-292 before pull-down. Representative result of 3 donors in 2 independent experiments is shown. *P < .05, 1-way ANOVA, followed by the Dunnett post hoc test.