Expanded Vδ2+ γδT cells were cocultured with live MV4-11, which had been precoated in magnetic anti-CD33 microbeads. After 30 min of coculture, MV4-11 was removed using magnetic column separation (Miltenyi, Bergisch Gladbach, Germany). The flow-through, containing effector cells only, was lysed and protein tyrosine kinase activity in the lysate was assessed using the PamChip PTK assay using a PamStation 12 (PamGene, Wolvenhoek, The Netherlands). Data were processed using BioNavigator6 (PamGene, Wolvenhoek, The Netherlands). A (normalized) kinase statistic Sk for the change in phosphorylation between samples in group x (target-free Vδ2) and group y (Vδ2 cocultured with MV-411) can be calculated as the mean signal-to-noise ratio of the individual peptides in the group: the mean signal in x minus the mean signal in y divided by the SD. If there is, on average, a larger change of the peptides in the same direction (i.e., all upwards or all downwards), a larger absolute value of Sk would result.

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