Journal: Nature Medicine
Article Title: IFN-γ-independent immune markers of Mycobacterium tuberculosis exposure
doi: 10.1038/s41591-019-0441-3
Figure Lengend Snippet: a , COMPASS analysis identified five functionally relevant CD4 T cell subset responses to Peptide Pool 2 (Ag85A, Ag85B and TB10.4), which are summarized in the heatmap. Rows represent study subjects and columns CD4 T cell functional subsets. The depth of shading within the heatmap represents the probability of detecting a response above background. IFN-γ-containing subsets are noted in red. b , Subject-specific COMPASS results in response to stimulation with Peptide Pool 2 were summarized using the polyfunctionality score, which weights T cell subsets that include more than one function. The total number of subjects analyzed was 41. Boxplots show median and interquartile ranges. The statistical significance was calculated using the Mann–Whitney U test, and the two-tailed P value is indicated. c , Representative flow cytometry plots from a ‘resister’ and an LTBI subject show frequencies of IFN-γ + CD40L/CD154 + IL-2 + TNF + T cells (red dots) in response to stimulation with Peptide Pool 2 or DMSO, with each experiment performed once. d , The absolute magnitude of Ag85/TB10.4-specific polyfunctional CD40L/CD154+ IL-2+ TNF+ CD4 T cells after background correction is displayed stratified by the expression of IFN-γ. These functional subsets represent the two right columns of the COMPASS plot in a . To facilitate visualization, we have not displayed a single LTBI outlier with the value of 3.41%. The total number of subjects analyzed was 41. Lines identify medians. The statistical significance was calculated using the Mann–Whitney U test, and two-tailed P values are indicated. e , COMPASS analysis identified 19 functionally relevant CD4 T cell subset responses to Mtb lysate, which are summarized in the heatmap. IFN-γ-containing subsets are noted in red. f , Polyfunctionality scores for the 41 subjects in response to Mtb lysate stimulation are shown, with boxplots representing median and interquartile ranges. The statistical significance was calculated using the Mann–Whitney U test, and the two-tailed P value is shown. g , Representative flow cytometry plots from a ‘resister’ and TST/IGRA-positive subject showing frequencies of IFN-γ + CD40L + IL-2 + TNF + T cells (red dots) in response to stimulation with Mtb lysate or DMSO are shown with each experiment performed once. h , The absolute magnitude of Mtb lysate-specific polyfunctional CD154 + IL-2 + TNF + CD4 T cells after background correction is displayed, stratified by the expression of IFN-γ for 40 subjects, with lines representing medians. These functional subsets represent the 9th and 17th columns of the COMPASS plot in e . To facilitate visualization, we have not displayed a single LTBI outlier with a value of 12.61%. The statistical significance was calculated using the Mann–Whitney U test, and unadjusted two-tailed P values are shown.
Article Snippet: Mtb antigens used were: PPD (Statens Serum Institute), recombinant Ag85A and -B in a 1:1 ratio (BEI Resources, NR-14871 and NR 14870), recombinant ESAT6 (BEI Resources, NR-14868) and CFP10 in a 1:1 ratio (BEI Resources, NR-49425), HspX (provided by T. Ottenhoff), GroES (provided by T. Ottenhoff) and LAM (BEI Resources, NR-14848).
Techniques: Functional Assay, MANN-WHITNEY, Two Tailed Test, Flow Cytometry, Expressing
