Figure 1.
Representative two-parameter dot plots of a patient with extrapulmonary TB (urogenital TB) indicating the frequency of PPD and ESAT-6-specific CD4+ T cells expressing IFN-γ and/or IL-2, respectively. PBMC were incubated with medium alone (control), PPD and ESAT-6, respectively.
Table 1.
Patient characteristics.
Figure 2.
Frequencies of ESAT-6 specific T cells.
Frequencies of single cytokine producing T cells and multifunctional T cells of 60 patients with pulmonary TB (light grey), 27 extrapulmonary TB (dark grey) and 91 with non-tuberculous diseases (white) after overnight stimulation with ESAT-6 are depicted. Boxes and whiskers are shown; the black line marks the median. Differences between patient groups are marked with a bar. A significantly increased frequency of CD4+ T cells expressing IFN-γ (0; 0–0; 0–0.07 vs. 0; 0–0.02; 0–0.3 or 0; 0–0.03; 0–0.3 [median; 25%–75% percentile; min–max], respectively. p = 0.034) and a decreased frequency of CD4+ T cells expressing IL-2 (0; 0–0.02; 0–0.26 vs. 0; 0–0.01; 0–0.06 [median; 25%–75% percentile; min–max]. p = 0.037) were found in patients with pulmonary TB when compared to diseases other then TB. CD8+ T cells expressing IFN-γ and TNF-α+ (0; 0–0.02; 0–0.16 vs. 0.02 ; 0–0.05; 0–0.23 [median; 25%–75% percentile; min–max]. p = 0.041) were different when pulmonary and extrapulmonary TB were compared. An independent-samples t-test was used to test for significance.
Figure 3.
Frequencies of PPD specific T cells.
Frequencies of single cytokine producing T cells and multifunctional T cells of 60 patients with pulmonary TB (light grey), 27 extrapulmonary TB (dark grey) and 91 with non-tuberculous diseases (white) after overnight stimulation with PPD are depicted. Boxes and whiskers are shown; the black line marks the median. CD4+IFN-γ+IL-10+ T cells (0.01 ; 0.02–0.33; 0–0.16 vs. 0; 0–0.01; 0–0.08 [median; 25%–75% percentile; min–max]. p = 0.007) were significantly different when extrapulmonary TB was compared to other diseases then TB. Comparing pulmonary and extrapulmonary TB, PPD specific CD8+ T cells expressing IL-2 (0.14 ; 0.09–0.365; 0–4.61 vs. 0.13; 0.06–0.33; 0–2.93 [median; 25%–75% percentile; min–max]. p = 0.011) were significantly different. Differences between patient groups are marked with a bar. An independent-samples t-test was used to test for significance.
Figure 4.
Receiver-operating-characteristic curves (ROC) were calculated for every significantly different cytokine and cytokine ratio. AUCs are summarized in Table 2.
Table 2.
Summary of receiver-operating-characteristic curves (ROC).
Figure 5.
Significant differences between different cytokine ratios are depicted, detected with a one way between-groups analysis of variance using the tukey test for post hoc analysis. Data from 60 patients with pulmonary TB (light grey), 27 extrapulmonary TB (dark grey) and 91 with non-tuberculous diseases (white) are depicted. Boxes and whiskers are displayed, the black line marks the median. Percentile ranges are additionally shown in Tables 3, 4, 5. (A) shows statistically significant differences between non-tuberculous diseases and pulmonary TB: PPD induced IFN-γ/IL-2 (p<0.001), ESAT-6 induced TNF-α/IFN-γ (p = 0.048), TNF-α/IL-2 (p = 0.03), IFN-γ/IL-2 (p = 0.005), all CD4+ T cell derived. (B) shows statistically significant differences between non-tuberculous diseases and extrapulmonary TB: PPD induced TNF-α/IL-2 (p<0.001), IFN-γ/IL-2 (p = 0.001), ESAT-6 induced TNF-α/IFN-γ (p = 0.026), TNF-α/IL-2 (p = 0.008), IFN-γ/IL-2 (p<0.001), CD4+ T cell derived. (C) shows statistically significant differences between pulmonary TB and extrapulmonary TB: PPD induced TNF-α/IL-2 (p = 0.001), CD4+ T cell derived.
Table 3.
Cytokine ratios pulmonary TB/non-tuberculous diseases.
Table 4.
Cytokine ratios extrapulmonary TB/non-tuberculous diseases.
Table 5.
Cytokine ratios pulmonary TB/extrapulmonary TB.