Analysis of renewal capacity of memory compartments and activated T cell replenishment of healed cutaneous leishmaniasis subjects.

<p>Renewal capacity of CD4⁺ (<b>A</b>) and CD8⁺ (<b>B</b>) T cells phenotipically characterized by central memory (Tcm, CD45RO⁺CCR7⁺) or effector memory (Tem, CD45RO⁺CCR7^-). The white box and whiskers represent those cells evaluated immediately after PBMC (<i>ex vivo</i>) and the gray boxes represent those cells after <i>in </i><i>vitro</i> stimuli with Lb-Ag. The central line represents median values. hCL= healed cutaneous leishmaniasis; HS= healthy subjects; *p<0.05, **p<0.01, ***p<0.001 (Kruskal-Wallis with post-test Dunns). Correlation analysis between the percentage of activated CD25⁺ (<b>C</b>, n=9) or CD69⁺ (<b>D</b>, n=12) in CD8⁺ T cells after <i>in </i><i>vitro</i> Lb-Ag stimuli and the percentage of effector memory (CD45RO⁺CCR7^-) CD8⁺ T cells after <i>in </i><i>vitro</i> Lb-Ag stimuli. Each point represents one subject. The graphs show the best fitted lines with 95% confidence intervals. r= correlation coefficient; p= significance level, (Spearman test).</p

Correlation analysis of clinical and/or immunological parameters from healed cutaneous leishmaniasis subjects.

<p>Correlation between the percentage of recently activated CD4⁺ (A, n=13) or CD8⁺ (B, n=14) T lymphocytes from PBMC after <i>in </i><i>vitro</i> Lb-Ag stimulation and the duration of clinical cure; Correlation between the percentage of activated CD4⁺ (C, n=8) or CD8⁺ (D, n=9) T lymphocytes and the concentration of IL-10 from cell culture supernatant; Each point represents one subject. The graphs show the best fitted lines with 95% confidence intervals. r= correlation coefficient; p= significance level, (Spearman test). </p

Cytokine mRNA expression in skin of hamsters immunised with <i>L. amazonensis</i> antigens (LaAg) and infected with <i>L. braziliensis</i>.

<p>Golden hamsters (6–7 per group) were immunised with IN LaAg (•), IM LaAg (▴) or PBS (□). Two weeks after the last immunisation, hamsters were challenged with <i>L. braziliensis</i>. (A) mRNA gene expression for IFN-γ and IL-10 in skin lesions was determined 114 days post-infection. (B) Correlation between lesion thickness and IFN-γ (r = 0.76, p<0.0001) or IL-10 (r = 0.68, p<0.0001). Each point represents one animal. The horizontal bars represent the median values. Symbols represent the fold change in cytokine expression relative to that of uninfected hamsters. Data shown are from one of two independent experiments.</p

Anti-<i>Leishmania</i> IgG and IgG2 serum levels in hamsters immunised with <i>L. amazonensis</i> antigens (LaAg) and infected with <i>L. braziliensis</i>.

<p>Golden hamsters (5–6 per group) were immunised with IN LaAg (•), IM LaAg (▴) or PBS (□). Two weeks after the last immunisation, hamsters were challenged with <i>L. braziliensis</i>. A) IgG and IgG2 serum levels were determined 114 days post-infection by ELISA. B) Correlation between lesion thickness and anti-leishmanial IgG (r = 0.78, p<0.0001) or IgG2 (r = 0.62, p<0.0001) serum levels. Each point represents one animal. The horizontal bars represent the median values, * p≤0.05. Data are from two independent experiments.</p

Parasite loads from skin lesions of hamsters immunised with crude <i>L. amazonensis</i> antigens (LaAg) and infected with <i>L. braziliensis</i>.

<p>Golden hamsters (4–6 per group) were immunised with IN LaAg (•), IM LaAg (▴), or received PBS (□). Two weeks after the last immunisation, hamsters were challenged with <i>L. braziliensis</i>. A) Paw skin parasite loads were determined 114 days post-infection via a limiting dilution assay. B) Correlation between skin parasite load and lesion thickness (r = 0.70, p<0.0001). Each point represents one animal. The horizontal bars represent the median values, * p≤0.05. Data are from two independent experiments.</p

Effectiveness of intranasal (IN) and intramuscular (IM) <i>L. amazonensis</i> antigens (LaAg) vaccination against subsequent infection with <i>L. braziliensis.</i>

<p>Golden hamsters (6–8 per group) were vaccinated with either 10 µg of LaAg intranasally (•) or 20 µg of LaAg intramuscularly (▴) followed by a booster vaccination 14 days later. Controls received PBS (□). Two weeks after the booster vaccination, animals were infected in the paw with 1×10⁵<i>L. braziliensis</i>. (A) Lesion thickness was scored on the indicated days. Means ± standard errors of the means (SEM), * p≤0.05, ** p≤0.01. (B) On day 114 post-infection, the lesion thickness of individual feet was shown as individual values, with the horizontal bars representing the median values. Data are from four independent experiments.</p