CD107 mobilisation assay

Day 10 Mϕs in 96-well tissue culture plates were washed three times in PBS and then cultured for 5 h with 1.52 × 10⁵ autologous Vδ2⁺ T cells per well in 200 μl complete medium to obtain an E:T ratio of 2:1 based on the initial seeding density of monocytes. Allophycocyanin-conjugated mouse anti-human CD107a (clone H4A3; Biolegend) and FITC-conjugated mouse anti-human CD107b (clone H4B4; Biolegend) or matched isotype controls were added directly to the wells at the start of the co-culture along with 1 μg/ml of monensin to neutralise intracellular acidity. Cells were then collected and labelled with PE-conjugated mouse anti-human Vδ2 (clone 123R3; Miltenyi Biotec) and PerCP-conjugated mouse anti-human CD3 (clone SK7; Biolegend) as described in “Flow cytometry”. Samples were acquired on an LSR II flow cytometer and analysed using FlowJo software. All comparatively analysed samples were acquired on the same day.