Group comparison of laboratory markers among total TB pleuritis patients with <50 (N = 41) and >50 (N = 88) CD4 counts.

*<p> = significant difference between the 2 groups.</p

Validity of ADA as a diagnostic test.

<p>PPV = positive predictive value; NPV = negative predictive value; CI = 95% confidence interval; LR+ = positive likelihood ratio; LR− = negative likelihood ratio.</p

Correlation (r) values for ADA and other blood and pleural fluid markers in tuberculous pleuritis patients (N = 197).

<p>LYM% = pleural fluid lymphocyte percentage of total leucocytes.</p

HIV status, CD4 counts and ADA values for the 2 groups with <50 (N = 25) and >50 (N = 33) CD4 counts among confirmed TB pleuritis patients.

<p>pos = positive, neg = negative, NA = not available, ADA = adenosine deaminase.</p

Effect of HIV antigen and MEK inhibition on FoxP3 expression in resting and activated Tregs in HIV patient samples.

<p>(a) Gating strategy for resting (CD4⁺CD45RA⁺FoxP3⁺) and activated (CD4⁺CD45RA-FoxP3⁺⁺) Tregs. (b) PBMCs were stimulated with <i>Gag</i> (grey boxes) or <i>Env</i> (white boxes) for 36 h in presence or absence of MEK inhibitor GSK1120212 or left untreated. Effect of GSK1120212 (100 nM and 10 μM) on FoxP3 expression levels (MFI) (c, e) and number of FoxP3⁺ T cells (d, f) in rTregs (c, d) and aTregs (e, f) stimulated with <i>Gag</i> or <i>Env</i>. (g) Ratio of aTreg over rTreg (%) in samples stimulated with <i>Gag</i> or <i>Env</i> alone and at two different MEKI concentrations as in d and e. (Boxes: median ± 25^th to 75^th percentile; whiskers: min to max, n = 8,* p< 0.05, **p <0.01).</p

Effect of MEK inhibition on CD4⁺ T cell cytokine expression in response to HIV antigen <i>Gag</i> and <i>Env</i>.

<p>Patient PBMCs were stimulated with <i>Gag</i> (grey boxes) or <i>Env</i> (white boxes) for 36 h and the percentages of (a) IFN-γ⁺, (b) TNF-α⁺, (c) IL-2⁺, (d) TNF-α⁺/IFN-γ⁺, (e) IFN-γ⁺/IL-2⁺, (f) TNF-α⁺/IL-2⁺ and (g) TNF-α⁺/IFN-γ⁺/IL-2⁺ cells were determined by intracellular staining and FACS analysis. (Boxes: median ± 25^th to 75^th percentile; whiskers: min to max, n = 8, * p< 0.05, ** p<0.01).</p

Effect of MEK inhibition on CD8⁺ T cell cytokine expression in response to HIV antigen <i>Gag</i> and Env.

<p>Patient PBMCs were stimulated with <i>Gag</i> (grey boxes) or <i>Env</i> (white boxes) for 36 h and the percentages of (a) IFN-γ⁺, (b) TNF-α⁺, (c) IL-2⁺, (d) TNF-α⁺/IFN-γ⁺, (e) IFN-γ⁺/IL-2⁺, (f) TNF-α⁺/IL-2⁺ and (g) TNF-α⁺/IFN-γ⁺/IL-2⁺ cells were determined by intracellular staining and FACS analysis.(Boxes: median ± 25^th to 75^th percentile; whiskers: min to max, n = 8, * p< 0.05,** p<0.01).</p

Effect of TB antigen and MEK inhibition on FoxP3 expression in resting and activated Tregs in TB patient samples.

<p>(a) Gating strategy for resting (CD4⁺CD45RA⁺FoxP3⁺) and activated (CD4⁺CD45RA⁻FoxP3⁺⁺) Tregs. (b) PBMCs were stimulated with ESAT-6/Ag85 for 36 h in presence or absence of 100 nM MEK inhibitor GSK1120212 or left untreated. Effect of GSK1120212 (100 nM and 10 μM) on FoxP3 expression levels (MFI) (c, e) and number of FoxP3⁺ cells (%) (d, f) in resting (c, d) and activated (e, f) CD4⁺Tregs stimulated with ESAT-6/Ag85. (g) Ratio of aTreg over rTreg (%) in samples stimulated with ESAT-6/Ag85 alone and at two different MEKI concentrations as in d and e. (Boxes: median ± 25^th to 75^th percentile; whiskers: min to max, n = 12, * p< 0.05, ** p<0.01, *** p< 0.001).</p

Effect of MEK inhibition on FoxP3 expression in sorted resting Tregs (rTregs).

<p>Human blood donor rTregs were stimulated with anti-CD3/CD28/CD2-coated MACSiBeads for 36 h in the presence or absence of MEK specific inhibitors (MEKI), followed by FoxP3 staining and FACS analysis. (a) Gating strategy for sorting of rTregs from CD4⁺ enriched cells. (b) rTregs were stimulated in the presence or absence of the MEK specific inhibitor GSK1120212 or left unstimulated. (c) rTregs were stimulated in presence or absence of the different MEK specific inhibitors FR180204, PD098059, U0126, CI-1040, AZD6244, PD0325901, MEK162, GSK1120212 at 1μM concentration. (d and e) Concentration-dependent effect of PD0325901 (IC₅₀ = 17 nM) and GSK1120212 (IC₅₀ = 4 nM). c-e: mean ± SD (n = 3).</p

Patient characteristics.

<p>^a) Lymphnode, pericardial, abdominal, cutaneous abscess, osteomyelitis.</p><p>^b) QuantiFERON-TB Gold^®.</p><p>^c) Erythrocyte Sedimentation Rate.</p><p>Patient characteristics.</p