Fig. 1.

RSTRs demonstrate IFN-γ independent T cell responses targeting ESAT-6 and CFP-10. (a) ICS data was generated from PBMCs from RSTR (n = 23) and LTBI (n = 23) subjects from the HETU study in response to stimulation with ESAT-6/CFP-10 peptide pool and analyzed by COMPASS. In the heatmap, rows represent study subjects, and columns represent CD4 T cell functional subsets. The depth of shading reflects the probability of detecting a response above background. CD4 subsets are indicated by bars at the bottom, with IFN-γ containing subsets shown in red. (b) Background corrected magnitudes (peptide pool minus DMSO) for each of the subsets identified by COMPASS are shown for RSTR (purple) and LTBI (green) individuals. Groups were compared by Mann–Whitney U test, and nominal p values < 0.05 are labeled. (c) Subject-specific COMPASS results were summarized for RSTR and LTBI individuals using the polyfunctionality score, which weights T cell subsets that include more than one function. Medians and interquartile ranges are depicted. The statistical significance was calculated using the Mann–Whitney U test, and the two-tailed p value is indicated. (d) Representative flow cytometry plots from two RSTR and one LTBI control subject are shown examining an IFN-γ-containing T cell subset in response to stimulation with DMSO or ESAT-6/CFP-10. Frequencies of T cells simultaneously producing IFN-γ, CD154, TNF, and IL-2 are shown and indicated as red dots in the typical two-dimensional layout. (e) Percentage of IFN-γ+ T cells for each subject was directly plotted against quantitative QFT result at baseline for LTBI subjects. (f) Percentage of IFN-γ+ T cells for each subject was directly plotted against quantitative QFT result at baseline for RSTRs with Spearman correlation coefficients (r) and p values indicated. (g) Percentage of IFN-γ+ T-cells was plotted against TST diameter at baseline for LTBI subjects. Spearman correlation coefficient and p value is indicated.