Increased frequency of Rv2628- and RD1-response in BALC than PBMC in active TB, evaluated by ELISPOT.

<p>ELISPOT evaluation of IFN-γ-producing CD4⁺ T-cells in circulating and BAL lymphocytes in response to Rv2628- and RD1-antigens in active TB and LTBI subjects. Response to Rv2628- (A) and RD1-antigens (B) in active TB patients; response to Rv2628- (C) and RD1-antigens (D) in LTBI subjects. <b>Footnote:</b> PBMC: peripheral blood mononuclear cells; BALC: bronchoalveolar lavage cells; SFC: spot-forming cells; IFN: interferon; TB: tuberculosis; LTBI: latent TB infection. Dotted lines link the results obtained for circulating and local lymphocytes for the same patient.</p

Increased frequency of Rv2628- and RD1-response in BALC than PB in active TB, evaluated by FACS.

<p>FACS analysis of the multiple cytokine producing CD4⁺ T-cells (IFN-γ and/or IL-2) in PB and BALC in response to Rv2628- (A) and RD1-antigens (B) stimulation in active TB patients. <b>Footnote:</b> PB: whole blood; BALC: bronchoalveolar lavage cells. Dotted lines link the results obtained for circulating and local lymphocytes for the same patient.</p

Cytokine profile in response to <i>Mtb</i>-specific antigens in active TB.

<p>Polyfunctional cytokine production analysis of Mtb-specific CD4⁺ T-cells by FACS. PB and BALC from active TB patients were stimulated overnight with Rv2628- and RD1-antigens. T- cells were classified as single IFN-γ-producing, single IL-2-producing or double IFN-γ/IL-2–producing cells in response to Rv2628- (A) and RD1-antigens (B). The results are reported as relative median in PB compared to BALC samples (A–B). Black triangles: PB; open triangles: BALC. <b>Footnote:</b> PB: peripheral blood; BALC: bronchoalveolar lavage cells; IFN: interferon; IL: interleukin.</p

Magnitude of RD1-response is significantly higher than Rv2628-response in PBMC and BALC, evaluated by ELISPOT.

<p>ELISPOT evaluation of IFN-γ-producing CD4⁺ T-cells in PBMC and BALC in response to Rv2628- and RD1-antigens in LTBI and active TB subjects. Response to Rv2628- and RD1-antigens in PBMC of active TB and LTBI subjects (A), response to Rv2628- and RD1-antigens in BALC of LTBI and active TB subjects (B) <b>Footnote:</b> PBMC: peripheral blood mononuclear cells; BALC: bronchoalveolar lavage cells; SFC: spot-forming cells; IFN: interferon; TB: tuberculosis; LTBI: latent TB infection.</p

Demographic and clinical characteristics of the subjects enrolled in the study.

<p><b>Footnotes:</b> TB: Tuberculosis; IQR: Interquartile range; BCG: Bacillus Calmette et Guérin; QTF-IT: QuantiFERON TB Gold in Tube; NA: not available.</p

Phenotype profile in response to <i>Mtb</i>-specific antigens in active TB.

<p>Phenotypical T-cell subset distribution of the total cytokine-producing T-cells in response to Rv2628- (A) or RD1-antigens (B) are reported in PB and BALC samples. <b>Footnote:</b> PB: peripheral blood; BALC: bronchoalveolar lavage cells; N: naïve; CM: central memory; EM: effector memory; EF: terminally^-differentiated effector memory.</p

BAL samples obtained and analyzed in the enrolled patients.

<p><b>Footnote:</b> ELISPOT: enzyme-linked immunosorbent spot; FACS: flow cytometric analysis; TB: tuberculosis; LTBI: latent tuberculosis infection; cell constraint: mononuclear cells from BAL < 6×10⁶; cell anergy: by ELISPOT, if the results are less than 20 SFC/well in the positive control; cell anergy: by FACS, if the results are less than 0.06% in the positive control; indeterminate: by ELISPOT, if the results are more than 10 SFC/well in the negative control.</p