The proportion of AgB-specific CD4⁺ T-cells producing IL-2⁺TNF-α⁺Th2⁺ (triple- positive) or TNF-α⁺Th2⁺ (double-positive) was increased in the “active stages” group compared to the “inactive stages” group.

<p>Proportion of the cytokine-producing subsets of the antigen-specific response in the different groups. <b>A.</b> Cytokine profile of the AgB-specific response in the “active stages” and “inactive stages” groups. <b>B.</b> Cytokine profile elicited by the control stimulus SEB in the “active stages”, “inactive stages”, and NO-CE subjects groups. The horizontal lines represent the median; statistical analysis was performed using the Mann-Whitney test, and p value was considered significant if ≤0.05.</p

Demographical and clinical characteristics of the enrolled subjects.

<p><b>Footnote</b>: N: Number; IQR: Interquartile Range; y: Year; US: Ultrasound.</p><p>Demographical and clinical characteristics of the enrolled subjects.</p

The proportion of triple-positive and double-positive T-cell subsets was increased in the “active stages” group compared to the “inactive stages” group.

<p>Proportion of the monofunctional and polyfunctional subsets in the “active stages” and “inactive stages” groups. The horizontal lines represent the median. Black dots indicate the “active stages” CE patients, white dots indicate the “inactive stages” CE patients. Statistical analysis was performed using the Mann Whitney test, and p value was considered significant if ≤0.05.</p

Flow chart of the enrolled subjects.

<p>Flow chart of the enrolled subjects.</p

Survey performed to enroll patients with suspected CE.

<p><b>Footnote:</b> CE: Cystic Echinococcosis</p><p>Survey performed to enroll patients with suspected CE.</p

Magnitude and cytokine profile of the total AgB-specific-response.

<p>Flow cytometry evaluation of CD4⁺ T-cell response to AgB. <b>A.</b> Magnitude of the cytokine response to AgB (considering the production of any cytokine) in the “active stages” and “inactive stages” groups. <b>B.</b> Frequency of the "total cytokine response" elicited by the AgB in the “active stages” and “inactive stages” groups. <b>C.</b> Frequency of the "total cytokine response" elicited by the control stimulus SEB in the “active stages”, “inactive stages” and NO-CE subjects groups. The positive CD4⁺ T-cell response was defined as the production of any cytokines (IFN-γ and/or IL-2 and/or TNF-α and/or Th2 cytokines and/or IL-10) with 0.03% as the detection limit corresponding to at least 30 analyzed events. The horizontal lines represent the median. Black dots indicate the “active stages” CE patients, white dots indicate the “inactive stages” CE patients. Statistical analysis was performed using the Mann-Whitney test, and p value was considered significant if ≤0.05.</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

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

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

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