Identification of activated B cells and antibody secreting cells in peripheral blood following Ty21a vaccination.

<p>PBMCs were collected from vaccinated or control (Ctrl) subjects at the indicated time points and analysis of different B cell populations was performed by flow cytometry. (A) Gating strategy for FACS analysis of activated B cells (ABC, blue) or antibody secreting cells (ASC, red). (B) Representative dot plots showing the proportions of ABC (CD20^hi CD71⁺) and ASC (CD20^- CD71⁺) in vaccinated and Ctrl individuals at the indicated time points. (C and D). Proportions of ABC and ASC in vaccinated and Ctrl individuals at the indicated time points. Dots represent individual values; bars indicate mean ± SEM. Statistical analysis was performed using one way ANOVA with Dunnett’s multiple comparison test for comparisons between individuals of the same group at different time points (pre- versus post-vaccination) (*, P< 0.05; **, P< 0.01).</p

Induction of anti-OmpC/F-specific porin antibodies in serum and stool after Ty21a vaccination.

<p>(A) IgM and (B) IgG, OmpC/F-specific antibody titers in serum samples from vaccinated subjects were analyzed by ELISA at the indicated time points after vaccination; participant numbers assigned according to increasing IgG reactivity. (C and D) Kinetics of anti-OmpC/F IgM (C) and IgG (D) antibody titers in vaccinated or control (Ctrl) individuals. Dots represent individual values; bars indicate mean ± SEM. (E) <i>S</i>. Typhi OmpC/F-specific IgA was determined from stool samples from vaccinated or Ctrl subjects by ELISA. Dots represent individual values; bars represent mean ± SEM. Statistical analysis was performed using one way ANOVA with Dunnett’s multiple comparison test for comparisons between individuals of the same group at different time points (pre- versus post-vaccination) (*, P< 0.05; **, P< 0.01 ***, P<0.001).</p

Activation profile of OmpC/F-specific CD4⁺ T cells after Ty21a vaccination.

<p>PBMCs were collected from vaccinated or control (Ctrl) subjects at the indicated time points. Activation of CD4⁺ T cells was assessed by flow cytometry after 24 h incubation with the indicated antigen. (A) Gating strategy for FACS analysis. (B) Representative dot plots showing the proportions of CD4⁺ CD40L⁺ T cells of Ctrl (upper panels) or vaccinated (lower panels) subjects after stimulation with <i>S</i>. Typhi OmpC/F porins or tetanus toxoid (TT). (C and D) Proportions of CD4⁺ CD40L⁺ T cells stimulated with TT (C) or OmpC/F (D) at the indicated time points. Dots represent individual values; bars represent mean ± SEM. (E) Representative dot plot showing IFN- γ and TNF-α production out of CD4⁺ CD40L⁺ cells after stimulation using medium or OmpC/F porins. (F) Proportions of OmpC/F-specific polyfunctional cells (IFN- γ ⁺ and/or TNF- α⁺) out of CD4⁺ CD40L⁺ T cells at indicated time points after vaccination in responder individuals. Statistical analysis in panels C and D was performed using one way ANOVA with Dunnett’s multiple comparison test for comparisons between individuals of the same group at different time points (pre- versus post-vaccination) or unpaired Student’s <i>t</i> test with Welch’s correction for comparison between Ctrl and vaccinated groups. Statistical analysis in panel F was performed using paired Student’s <i>t</i> test for comparison between individuals of the same group (Day 11 versus Day 60) (*, P< 0.05; **, P< 0.01).</p

ROCs of transient elastography, APRI, FIB-4, Fibrotest for cirrhosis (F4).

<p>ROC: receiver operating characteristic curve; fibro: transient elastography (TE); fibtest: Fibrotest®.</p

Standard cut-off values of non-invasive tests of liver fibrosis in the literature.

<p>Standard cut-off values of non-invasive tests of liver fibrosis in the literature.</p

Performance of non-invasive tests for diagnosis cirrhosis (F4).

<p>TE: transient elastography; Hya: hyaluronic acid, ELF: Enhanced Liver Fibrosis-Test; Sens: sensitivity, Spec: specificity, PPV: positive predictive value, NPV: negative predictive value</p><p>Performance of non-invasive tests for diagnosis cirrhosis (F4).</p

ROCs of transient elastography, hyaluronic acid, Hepascore, ELF-Test for significant fibrosis (F> = 2).

<p>ROC: receiver operating characteristic curve; fibro: transient elastography (TE); hyaluron: hyaluronic acid; hepasco: Hepascore, ELF: Enhanced Liver Fibrosis-Test®.</p

ROCs of transient elastography, hyaluronic acid, Hepascore, ELF-Test for cirrhosis (F4).

<p>ROC: receiver operating characteristic curve; fibro: transient elastography (TE); hyaluron: hyaluronic acid; hepasco: Hepascore, ELF: Enhanced Liver Fibrosis-Test®.</p

ROCs of transient elastography, APRI, FIB-4, Fibrotest for significant fibrosis (F> = 2).

<p>ROC: receiver operating characteristic curve; fibro: transient elastography (TE); fibtest: Fibrotest®.</p

Blood markers for non-invasive diagnosis of liver fibrosis.

<p>Blood markers for non-invasive diagnosis of liver fibrosis.</p