<i>Ex vivo</i> activation of T cells.

Fresh tonsil tissue slices were incubated with or without stimulant (PHA-L/IL-2) and either with or without the presence of the inhibitor cocktail. All slices were incubated for 96 h. A. Slices were harvested and mechanically disrupted before staining for flow cytometry. Plots were gated for CD3, CD4, or CD8 and Ki67, with HLA-DR as a counterstain. Percentage of CD3+CD4+ or CD3+CD8+ T cells that are Ki67+ is shown. Figure depicts 1 tonsil donor, representative of N = 10 tonsil donors from at least 5 independent experiments. B. Fold-change analysis of T-cell activation (Ki67) from tissue slices analysed via flow cytometry. Y axis represents fold change from stimulated controls. A 2-tailed Wilcoxon signed rank test was used to obtain P-values comparing activated+inhibitor to activated; ** P N = 10 tonsil donors from at least 5 independent experiments. C. Confocal microscopy of tonsil tissue slices stimulated with PHA-L + IL-2 for 90 h with or without inhibitor cocktail. CD3 in green, Ki67 in white. Scale bar in white represents 50 μm. Representative example of 2 independent experiments. Data used in the generation of this figure can be found in S1 Data. PHA-L/IL-2, phytohaemagglutinin-L/interleukin 2.