Parasitological evaluation of placebo and vaccinated dogs.

<p>The presence of (A) live <i>Leishmania</i> parasites was highlighted by subculture analysis of bone marrow aspirates isolated from dogs of placebo (n = 5), rPSA/QA-21 (n = 9) and Cter-rPSA (n = 5) groups at 2, 4 and 6 months post-challenge (PC). A sample was considered as positive when <i>Leishmania</i> parasites were detected during the seeding or subculture analysis. The presence of (B) <i>Leishmania</i> DNA and (C) the parasite load in bone marrow aspirates of dogs of each group were assessed by quantitative PCR. Dogs were considered as positive when the titer was superior to 40 parasites per mL. Data are expressed as (B) the number of dogs with positive PCR at each time points post-challenge and (C) the mean number of parasites per mL of bone marrow aspirates at different times post-challenge (4 and 6 months) (* <i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001).</p

Vaccine specific serological responses as detected by Enzyme-Like Immunosorbent Assay (ELISA).

<p>Evolution of levels of (A) anti-rPSA, (B) anti-Cter-rPSA and (C) anti-<i>Li</i>ESAp specific IgG2 antibodies was assessed in serum samples isolated from dogs of each group immediately before immunization and at different times post-immunization: first month after the second dose and two months after the third dose. Each serum sample was tested in triplicates. Cut-off value was calculated using the following formula: mean OD in sera from all dogs before immunization + 3 standard deviations. Values represent means OD +/- standard deviation of triplicate experiments (* <i>p</i>< 0.05, ** <i>p</i><0.01, *** <i>p</i><0.001).</p

Anti-leishmanial activity of canine monocyte-derived macrophages in non-immune and immune dogs.

<p>The ability of pre-infected canine monocyte-derived macrophages to kill <i>Leishmania</i> parasites when they were exposed to autologous peripheral lymphocytes derived from PBMC was expressed as the percentage of parasitic index inhibition after <i>in vitro</i> infection with <i>Leishmania infantum</i> promastigotes (MHOM/MA/67/ITMAP-263) and 72 h incubation with and without autologous lymphocytes. Anti-leishmanial activity of co-cultured canine macrophages was evaluated immediately before immunization and two months after the third dose. Values represent means +/- standard deviation of duplicate experiments (* <i>p</i>< 0.05, ** <i>p</i><0.01, *** <i>p</i><0.001).</p

Cytokine levels measured in the CSF and plasma of HAT patients according to the disease stage.

1<p>Interquartile range.</p>2<p>P values are given for the three class comparisons (stage1, early stage 2, and late stage 2), p-values<0.05 are in bold.</p><p>Cytokine levels measured in the CSF and plasma of HAT patients according to the disease stage.</p

Cytokine profiles and infection outcomes in gambiense HAT.

<p>Cytokine profiles and infection outcomes in gambiense HAT.</p

Plasmatic cytokine profiles in HAT patients, SERO TL+ subjects, and endemic controls.

<p>Box plots of cytokine concentration are presented. Boxes represent the medians and interquartile ranges and whiskers represent 10^th and 90^th percentiles. Asterisks indicate significant differences over the control group: ****, p<0.0001; ***, p<0.001; **, p<0.01; *, p<0.05 (nonparametric Wilcoxon signed-rank test). Only cytokines with significant differences are shown. Group effectives: control (n = 40); HAT (n = 52); SERO TL+ (n = 60).</p

Prognostic value of cytokine levels in SERO TL+.

<p>The prognostic value of cytokine levels measured at inclusion was evaluated by stepwise multivariate logistic regression analysis. For this, SERO TL+ individuals were spited into three categories according to follow-up results: those who were confirmed as HAT patients (SERO TL+/HAT; n = 12; G1), those who presented decreasing CATT responses (SERO TL+/CATTneg; n = 15; G2) and those who maintained high CATT responses and remained negative in parasitology (SERO TL+/CATT≥1/8; n = 13; G3). Three models were then constructed independently by a stepwise procedure comparing SERO TL+/HAT (model I), SERO TL+/CATTneg (model II) and SERO TL+/CATT≥1/8 (model III) to the two other groups.</p>1<p>The covariate univariate p-value before the stepwise procedure is shown in between brackets.</p>2<p>Cytokine levels were assigned to two class of equal size using the cytokine median value as the threshold.</p>3<p>Odds ratio.</p>4<p>Confidence interval.</p><p>Prognostic value of cytokine levels in SERO TL+.</p

Principal component analysis of cytokine responses in HAT patients and SERO TL+.

<p>A. Histogram of eigenvalues. The eigenvalues, which corresponds to the amount of variance accounted by each component, are presented for the first ten components. The two first components, which explain more than 50% of the total variance, are in black. B. First factorial plan (x-axis: first component, y-axis: second component). The plan composed by the two first axis or dimensions is represented. It corresponds to an approximation of the cloud of points by a 2-dimensional space. C. Absolute contribution of each cytokine to the variance of the second component.</p

Phenotypic and demographic characteristics of the study sample.

<p>Phenotypic and demographic characteristics of the study sample.</p

Total IgG prevalence against <i>La</i>PSA-38S in different endemic areas of leishmaniasis.

<p>The cut-off value of reactivity for <i>La</i>PSA-38S antigen was calculated as the mean plus 3 SD of the OD values observed in naive controls from each country. These cut-off values allowed us to distinguish between positive and negative sera and consequently to estimate performance parameters of each ELISA test.</p