Treg frequency and function increase after treatment of CDL.

<p>Frequency of cells with a Treg phenotype in PBMCs and suppressive capacity of CD4⁺CD25⁺ cells were determined for CDL patients (n = 11) before and after treatment. <b>A</b>. Frequency of CD4⁺CD25^hiFoxp3⁺ cells. <b>B</b>. Frequency of CD4⁺CD25^hiCD127⁻ cells. <b>C</b>. Frequency of CD4⁺CD25^hiGITR⁺ cells. <b>D</b>. Inhibition of CD4⁺CD25⁻ cell proliferation by CD4⁺CD25⁺ cells at a 1∶1 ratio after stimulation with <i>L. panamensis (L.p.)</i> and APCs. <b>E</b>. Inhibition of CD4⁺CD25⁻ cell proliferation by CD4⁺CD25⁺ cells at a 4∶1 ratio after stimulation with PHA. <b>F</b>. Inhibition of CD4⁺CD25⁻ cell IFN-γ secretion by CD4⁺CD25⁺ cells at a 1∶1 ratio after stimulation with <i>L.p.</i> and APCs. Inhibition was calculated only for subjects that had proliferation and IFN-γ levels above the average+2SD of control wells from the same co-culture (n = 10 for proliferation and 11 for IFN-γ secretion with <i>L.p.</i> stimulation and n = 11 for PHA stimulation). *p<0.05, paired t-test, **p<0.01, Wilcoxon signed-rank test.</p

IL-10 does not mediate suppression of effector functions by CD4⁺CD25⁺ cells.

<p>IL-10 was measured in supernatants from the CD4⁺CD25⁻-CD4⁺CD25⁺ co-cultures by ELISA. The level of IL-10 in the absence (1∶0 ratio) or presence (1∶1 ratio) of CD4⁺CD25⁺ cells is shown for AI (n = 6), CDL patients before treatment (n = 14), CDL patients after treatment (n = 11) and all co-cultures combined (n = 31). **p<0.01, Wilcoxon signed-rank test. Means with SEM are shown.</p

CDL patients have a higher frequency of CD4⁺CD25^hiFoxP3^<b>+</b> cells in peripheral blood than AI.

<p>PBMCs from AI (n = 12) and CDL patients (n = 14) were stained for CD4, CD25 and either Foxp3, CD127, or GITR and analyzed by flow cytometry. <b>A</b>. Frequency of CD4⁺CD25^hiFoxp3⁺ cells. <b>B</b>. Frequency of CD4⁺CD25^hiCD127⁻ cells. <b>C</b>. Frequency of CD4⁺CD25^hiGITR⁺ cells. ***p<0.001, Mann-Whitney test. The median is represented by a horizontal line.</p

Relative expression of <i>IDO</i> decreases in CDL patients after treatment and is correlated with <i>IFNG</i>.

<p>RNA was isolated from biopsies of lesions from CDL patients (n = 11) before and after treatment and the expression of <i>FOXP3</i> (<b>A</b>), <i>IFNG</i> (<b>B</b>), <i>IL10</i> (<b>C</b>) and <i>IDO</i> (<b>D</b>) was measured by qRT-PCR. The expression of each gene relative to healthy skin of four normal controls was calculated using the ΔΔCT method. <b>E</b>. Correlation between the relative expression of <i>IDO</i> and <i>FOXP3</i> (left panel) or <i>IFNG</i> (right panel). *p<0.05, Wilcoxon signed-rank test.</p

Clinical characteristics and evolution of chronic dermal leishmaniasis patients.

a<p>% of surface area of ulcer(s) or plaque(s) healed or volume of nodule(s) reduced at the end of treatment; ND: not determined because culture was not necessary or no strain was obtained; NA: not applicable.</p