Figure 5.
Effects of BTKi’s on platelet ATP secretion, P-selectin expression, and platelet-neutrophil complex formation after FcγRIIA stimulation by cross-linking. Blood was incubated for 30 minutes with solvent (DMSO, 0.1%) or BTKi’s (ibrutinib [ibr], 0.2 µM; acalabrutinib [aca], 1 µM; tirabrutinib [tira], 1 µM; zanubrutinib [zanu], 0.4 µM; and evobrutinib [evo], 2.5 µM) and for 3 minutes with AT10 (2 µg/mL) before stimulation with Fab2 (30 µg/mL). (A) ATP secretion was measured by using the LUMI-aggregometer. Values are mean ± SD (n = 5). (B) Platelet P-selectin expression (measured by flow cytometry) analyzing the mean fluorescence intensity (MFI) of P-selectin. (C) Platelet-neutrophil complexes determined by flow cytometry as the mean fluorescence intensity of CD41 (platelets) on CD66b cells (neutrophils). (B-C) Scatter plots from 3 experiments with different blood donors. *P < .05.