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. 2019 Jul 3;10:1518. doi: 10.3389/fimmu.2019.01518

Figure 2.

Figure 2

IFN-γ positive CD4+ T-cell proportions specific for E6/C10 and latency Mtb antigens after short-term and two-hit in vitro culture. IFN-γ positive CD4+ T cells of TB patients and HCs were measured by intracellular flow cytometry after stimulation short-term (A,B), long-term culture (two-hit) (C,D) or comparison between both (E,F). E6/C10, PPD, or PHA were used in short-term whole blood (A) or purified PBMCs culture (B). E6/C10, PPD, and latency antigens (i.e., E6/C10, Rv1733, Rv2628, Rv2031, or Rv3407) were used in long-term two-hit stimulation C,D). Symbols indicate mean values of duplicates for individual tuberculosis patients (black triangles) and HCs (open circles). (E,F) Paired comparison of short-termE6/C10 specific CD4+ T-cell proportions with long-term two-hit stimulation for E6/C10 (E) and Rv1733, Rv2628 (F) in TB patients (left graphs) and HCs (right graphs). Significant differences are indicated by asterisks. Nominal p-values for the Mann-Whitney U-test (two-tailed) were calculated and shown as *p < 0.05; ***p < 0.001 and ****p < 0.0001. ns, not significant.