IFN-γ/IL-17A/IL-22 triple FluoroSpot analysis of PBMC responses to CA, TT and PPD. PBMC incubated with stimuli or medium alone were analyzed for spots representing cells secreting single, double and triple combinations of IFN-γ, IL-17A and IL-22. (A) The bars represent individual responses (n = 8) to all antigens divided into single, double or triple spots; (B) Analysis of the proportion of double- and triple-stained spots induced by the different antigens shown for all eight donors. The box plot show median (line in box), min and max (vertical lines), and 25 and 75 percentiles (box) for CA, TT, and PPD stimulation (n = 8). Statistical significance is indicated as * (p < 0.01) or ns (not significant); (C) Average proportion of single and double spots for each cytokine/cytokine combination per total number of spots. PBMC with responses below 40 spots in total were excluded for the analysis (1 donor for CA, 2 donors for TT and 1 donor for PPD) to avoid an impact of subtle differences in spot numbers in the low responders.