Fig. 4: Characterisation of Vγ9Vδ2 T cell serial killing dynamics.

a–d Measured killing of the A498GBDR targets by γδD4 T cells +/− 1.5 μM HMBPP in 350 μm-well chip during 16 h of imaging each position every 3 min. a Percentage of γδ T cell that kill at least one target n = 6 donors and b percentage of γδ T cell killers that killed more than one target cell, n = 6 donors. The box limits represent upper and lower quartiles, and whiskers indicate the minimum and maximum values. One dot represents the measurement for one donor. c, d Proportion of serial killing (Pie chart) and killing strategies used during serial killing aggregated for all γδD4 T cells (top bar graph) or for individual γδD4 T cells (lower diagram). Numbers above each bar of the top bar graph represent the respective number of events. Results from 6 donors were pooled. c Control condition and d HMBPP-treated cells. e Proportions of killing strategies comparing γδD4 T cells killing a single target cell with the first target killed from γδD4 T serial killers (data from c, d). f Number of killed target cells per γδD4 T cell (targets treated with 1.5 μm HMBPP) or NKD4 cell killers for each killing mechanism, n = 4 donors. g In-tube 4 h degranulation assay of γδD4 T cells against wild-type A498 measured by flow cytometry at the indicated timepoints. Surface expression of CD95/FasL (n = 3 donors) and TRAIL (n = 4 donors) showed after gating on degranulating (CD107a+) γδ T cells. Control is unstained. All applicable plots show means with standard deviations unless specifically stated otherwise. Statistics were calculated using two-tailed paired t-test (a, b, f) and one-way ANOVA (g). ns=not-significant.