Heatmap summarizing the spot intensities of the peptide microarray for the selected peptides.

Microarray reactivities to selected peptides by testing sera from the screening cohort, comprising 50 patients with CE and 25 controls with non-parasitic focal liver lesions. The heatmap depicts the distribution of the spot intensities from the peptide microarray according to the antigenic reaction. The blue and red colors indicate low and high antigenic response respectively. The clustering is generated from the heatmap and it groups the patients (y-axis) and the peptides (x-axis) according to the antigenic response from the microarray. Footnotes: CE, cystic echinococcosis.</p

Heatmaps representing the spot intensities of peptide microarray for the selected peptides comparing CE patients and controls or active and inactive CE cysts.

Left panel. Spot intensities were analyzed comparing CE patients (n = 50) and controls (n = 25) from the screening cohort. Right panel. Spot intensities were analyzed within CE patients only comparing patients with active cysts (n = 30) and patients with inactive CE cysts (n = 20) from the screening cohort. Footnotes: CE, cystic echinococcosis.</p

Background analysis of CE and controls sera.

Analysis of the background using a home-made ELISA by testing sera from the validation cohort comprising 29 patients with cystic echinococcosis and 14 controls with non-parasitic focal liver lesions. The optical density (OD) read for the wells where no peptides were adsorbed (no Ag) (“peptide-/serum+/secondary antibody+”) was considered as the background and compared with the OD read where peptides were adsorbed (Ag) (“peptide+/serum+/secondary antibody+”). Horizontal lines represent medians. P value was considered significant if (DOCX)</p

Demographical and clinical characteristics of the screening and validation cohorts.

Demographical and clinical characteristics of the screening and validation cohorts.</p

Sensitivity and specificity of the candidate antigens identified through peptide microarray.

Sensitivity and specificity of the candidate antigens identified through peptide microarray.</p

Bioinformatics and experimental workflow to identify most exposed proteins and immunogenic peptides starting from the <i>E</i>. <i>granulosus</i> s.s. genome.

Starting from all proteins predicted from the genome, TMHMM2, SignalP4 and predGPI software identified the most exposed proteins of E. granulosus. The localization of these proteins was further evaluated using the DeepLoc software and all proteins were retained for the second step of the pipeline to determine specific B-cell antigenic epitopes through Bepipred, SVMTrip e LBTOPE software. All peptides were filtered to avoid redundancy. Finally, peptides were screened according to the hydrophilicity characteristics using the Parker’s algorithm.</p

None of the peptides associate to specific CE cyst stage.

Validation of microarray results using a home-made ELISA by testing sera from the validation cohort comprising 29 patients with cystic echinococcosis and 14 controls with non-parasitic focal liver lesions. Peptides resulting from the manual examination of the microarray results promising for the diagnosis of CE infection (EGR_08002; EGR_03286; EGR_03099; EGR_10786) or for discriminating between active and inactive CE cysts are analyzed comparing CE patients with cysts at different stages. The following groups are compared: active cysts (CE1 and CE2/3b), transitional cysts (CE3a), inactive cysts (CE4/CE5 no therapy), and inactive cysts that received therapy in the last 5 years (t (DOCX)</p

Flow chart of the enrolled subjects.

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