Phagocytosis and the killing of <i>L</i>. <i>braziliensis</i> by monocytes from CL patients.

<p>Monocytes from CL patients (n = 9) and HS individuals (n = 6) were infected with <i>L</i>. <i>braziliensis</i> promastigotes at a 5:1 ratio for 2, 24, 48 and 72 hours. The number of infected cells (A) and the number of intracellular parasites (B) were determined by microscopic evaluation after May-Grunwald-Giemsa staining from cytocentrifuge preparations. Monocytes were preincubated with either DPI (10mM) or L-NMMA (1mM), for 10 minutes and were infected with <i>L</i>. <i>braziliensis</i> promastigotes at a 5:1 ratio for 72 hours. (C) The number of infected cells. (D) The number of intracellular parasites. Statistical analysis was performed using the Kruskal-Wallis test (* p < 0.05, ** p < 0.01).</p

IL-22 does not alter the immune response during <i>L</i>. <i>major</i> infection.

<p>Wild-type and <i>Il22</i>^-/- mice were intradermally infected with 2 x10⁶<i>L</i>. <i>major</i> promastigote metacyclics and cells from 5 week old lesions were collected and analyzed by flow cytometry. <b>(A)</b> Representative dot plots and bar graphs depict frequencies and total cell numbers of CD4+, CD8+, CD11b+, and LY6G+ cells. <b>(B-C)</b> RNA was isolated from the lesions of wild-type mice infected with <i>L</i>. <i>major</i> to assess gene expression. Data are represented as relative expression to housekeeping gene <i>rps11</i> and are representative of at least 3 independent experiments, with 3–5 mice per group. Error bars indicate mean ± SEM, *p < 0.05,*p < 0.01, ***p < 0.001.</p

Production of CXCL9, CXCL10 and CCL5 by macrophages from HTLV-1 infected subjects.

<p>Macrophages from healthy subjects (HS), HTLV-1 carriers (HC) and from HAM/TSP patients were cultured without stimulus or with LPS for 48 hours to evaluate (A) CXCL9 and (B) CXCL10 and (C) CCL5 productions. ELISA was used to measure these chemokines levels. Data is represented by median and range. Kruskal-Wallis followed by Dunn's post test (* <i>P</i><0.05) and Wilcoxon T test (# <i>P</i><0.05) were used for statistical analyses.</p

Correlation between IFN-γ produced by PBMCs and intermediate monocytes frequency from HAM/TSP patients.

<p>Correlation between IFN-γ production by PBMCs and intermediate monocytes frequency from HAM/TSP patients (n = 12). Spearman correlation and r significance test were used for statistical analyses (<i>P</i><0.05).</p

Percentage of infected macrophages and number of amastigotes/100 macrophages from HTLV-1 infected subjects.

<p>Macrophages from healthy subjects (HS, n = 10), from HTLV-1 carriers (HC, n = 10) and from HAM/TSP patients (n = 10) were infected by <i>L. braziliensis</i> at stationary phase at 5∶1 ratio. (A) Percentage of infected macrophages and (B) the number of amastigotes/100 macrophages were evaluated after 2, 48 and 72 hour of infection. Data is represented by median. Kruskal-Wallis followed by Dunn's post test were used for statistical analyses (*<i>P</i><0.05).</p

Oxidative burst production before and after therapy and cure of CL patients.

<p>Production of burst oxidative, NO and ROS by monocytes from CL patients (n = 6) after infection with <i>L</i>.<i>braziliensis</i> promastigotes or upon PMA stimulus, were determined before and after therapy (i.v. pentavalent antimonial, 20mg/kg body weight daily for 20 days) and cure of cutaneous leishmaniasis. The data represent the median of mean intensity of fluorescence (MIF) of oxidative burst production (A), frequency of NO production (B) and frequency of ROS production (C). Statistical analysis was performed using Wilcoxon test and results were considered significant (p<0.05).</p

Clinical and epidemiological characteristics of cutaneous leishmaniasis patients (CL) and Healthy Subjects (HS).

<p>Clinical and epidemiological characteristics of cutaneous leishmaniasis patients (CL) and Healthy Subjects (HS).</p

IL-22 regulates the expression of skin repair genes during <i>L</i>. <i>major</i> infection.

<p><b>(A-B)</b> Wild-type and <i>Il22</i>^-/- mice were intradermally infected with 2 x10⁶<i>L</i>. <i>major</i> promastigote metacyclics and RNA was isolated from the lesions at 5 weeks post-infection to assess gene expression. Data are represented as relative expression to housekeeping gene <i>rps11</i> and are representative of at least 2 independent experiments, with 3–5 mice per group. Error bars indicate mean ± SEM, *p < 0.05.</p

Production of TNF and IL-10 by macrophages from HTLV-1 infected subjects.

<p>Macrophages from healthy subjects (HS), HTLV-1 carriers (HC) and from HAM/TSP patients were cultured without stimulus, or with LPS for 48 hours to evaluate (A) TNF and (B) IL-10 productions. ELISA was used to measure these cytokines levels. Data is represented by median and range. Kruskal-Wallis followed by Dunn's post test (* <i>P</i><0.05) and Wilcoxon T test (# <i>P</i><0.05) were used for statistical analyses.</p

Monocytes from CL patients produced high levels of reactive oxygen species after infection with <i>L</i>.<i>braziliensis</i>.

<p>Monocytes from CL patients (n = 13) and HS individuals (n = 7) were stained with DAF-FM diacetate (NO probe, 10mM) and CMH-2DCFDA (ROS probe, 1 μM) for 10 minutes, infected with <i>L</i>.<i>braziliensis</i> promastigotes for 25 minutes at a ratio of 5:1cells, and stained with anti-CD14. PMA was used as positive control. Data were collected using flow cytometry and analyzed using FLOWJO software (A). Representative histograms of ROS production, (B) Frequency of <i>L</i>.<i>braziliensis</i>-infected monocytes expressing ROS, (C) Representative histograms of NO production, (D) Frequency of <i>L</i>.<i>braziliensis</i>-infected monocytes expressing ROS. Statistical analysis was performing using ANOVA with Bonferoni´s pos-test and Manny Whitney test. The results were considered significant with a p< 0.05 (*p<0.05).</p