Fig. 2: H4:IC31-induced immune responses.

tSNE analysis of total cytokine-expressing lymphocytes observed 70 days post-vaccination in H4:IC31 recipients (n = 26) after in vitro restimulation of whole blood with Ag85B (a) or TB10.4 (b). Mean proportions of each lymphocyte subset among the total cytokine-expressing cells are indicated by the horizontal bars. Error bars represent the 95% CI and colors correspond to those in the tSNE plot. Three cell populations were not identifiable with the flow cytometry antibody panel. c Frequencies of CD4 T cells expressing any combination of IFNγ, IL-2, TNF, IL-22, and/or IL-17 for each individual at day 0 (circles) and day 70 (diamonds) randomized to placebo (blue) or H4:IC31 (red). Values above the horizontal floating lines represent p values obtained by comparing responses between day 0 and day 70, calculated by Wilcoxon signed-rank test. d Relative proportions of total cytokine-expressing Ag85B- (blue) or TB10.4-specific (orange) CD4 T cells expressing Th1 cytokines (IFNγ, IL-2 and/or TNF), IL-22 or IL-17 measured at day 70 in H4:IC31 recipients. All participants had a detectable response (Fishers’ exact test, see “Methods”). e, f COMPASS polyfunctionality scores for CD4 T cells, stratified by vaccine arm in response to Ag85B (e) and TB10.4 (f). Changes between day 0 (circles) and day 70 (diamonds) were calculated by Wilcoxon signed-rank test. g Heatmap of COMPASS posterior probabilities for detecting an antigen-specific CD4 T cell response on day 70 relative to day 0 for the indicated cytokine-co-expression subsets in H4:IC31 or placebo recipients. Columns correspond to the different cell subsets (shown are the 13 of 24 subsets with detectable antigen-specific response in at least one participant regardless of antigen specificity), identified below the heatmap and color-coded by the cytokines they express (white = none, shaded = present) and ordered by degree of functionality from one function on the left to five functions on the right. Each row in the heatmap represents one participant. h Percentage of participants with significant vaccine-induced responses to Ag85B (blue) or TB10.4 (red), calculated by MIMOSA2 (see “Methods” section), based on CD4 T cells expressing any combination of IFNγ, IL-2, TNF, IL-22, and/or IL-17 (left), polyfunctional IFNγ+IL-2+ TNF+ (center) or Th22 (right) cytokines on day 70 relative to day 0.