Upregulated events related to cell-to-cell signaling and interaction, immune cell trafficking and inflammatory response.

<p>Ingenuity Pathway Analysis. The most important functional categories participating in the three major biological activities selected for localized cutaneous leishmaniasis (LCL, <i>n</i> = 5) samples are listed above for both LCL and mucosal leishmaniasis (ML, <i>n</i> = 5) groups. Only events with <i>P</i>-values higher than 0.001 were included in this list. <i># Genes</i> indicates the number of genes from each list that were involved in the events.</p

Global transcriptional expression patterns.

<p>Hierarchical clustering was performed using the Euclidean distance method (Gene Cluster 3.0) with complete linkage for both samples and genes assays. Intensity of gene expression increases from black to yellow. The heat map in the left panel includes the 18,577 genes detected by RNA-seq and shows that the samples were correctly clustered into the two clinical phenotypic groups, ML and LCL. The right panel represents the same assay using the 1208 genes selected by the dimensional reduction step performed by the <i>t-test</i>, confirming that the samples could still be clustered into LCL and ML groups. LCL: Localized Cutaneous Leishmaniasis. ML: Mucosal Leishmaniasis.</p

Genes associated with high risk for mucosal leishmaniasis progression.

<p>Genes used in Leave-One-Out cross validation analysis or selected by Ingenuity Pathway Analysis biomarker filter tools. Possible biomarkers to predict the development of mucosal leishmaniasis were determined by IPA analysis and have a significant fold change and a predicted importance due to their use as diagnostic and prognostic biomarkers in other diseases.</p><p>FC: fold change. NE: No expressed.</p

Transcriptome Patterns from Primary Cutaneous <em>Leishmania braziliensis</em> Infections Associate with Eventual Development of Mucosal Disease in Humans

<div><h3>Introduction</h3><p>Localized Cutaneous Leishmaniasis (LCL) and Mucosal Leishmaniasis (ML) are two extreme clinical forms of American Tegumentary Leishmaniasis that usually begin as solitary primary cutaneous lesions. Host and parasite factors that influence the progression of LCL to ML are not completely understood. In this manuscript, we compare the gene expression profiles of primary cutaneous lesions from patients who eventually developed ML to those that did not.</p> <h3>Methods</h3><p>Using RNA-seq, we analyzed both the human and <em>Leishmania</em> transcriptomes in primary cutaneous lesions.</p> <h3>Results</h3><p>Limited number of reads mapping to <em>Leishmania</em> transcripts were obtained. For human transcripts, compared to ML patients, lesions from LCL patients displayed a general multi-polarization of the adaptive immune response and showed up-regulation of genes involved in chemoattraction of innate immune cells and in antigen presentation. We also identified a potential transcriptional signature in the primary lesions that may predict long-term disease outcome.</p> <h3>Conclusions</h3><p>We were able to simultaneously sequence both human and <em>Leishmania</em> mRNA transcripts in primary cutaneous leishmaniasis lesions. Our results suggest an intrinsic difference in the immune capacity of LCL and ML patients. The findings correlate the complete cure of <em>L. braziliensis</em> infection with a controlled inflammatory response and a balanced activation of innate and adaptive immunity.</p> </div

Genes involved in the most relevant biological activities according to Ingenuity Pathway Assay.

<p>The bars indicate the significance levels of the up-regulated genes presence based on the normal event (yellow line). Black and gray bars represent LCL and ML samples, respectively. LCL: Localized Cutaneous Leishmaniasis. ML: Mucosal Leishmaniasis.</p

Top 10 biological pathways observed in American tegumentary leishmaniasis lesions based on clinical presentation.

<p>Assay using Ingenuity Pathway Analysis for gene classification according to functional annotation. Biological activities were ordered according to <i>P</i>-values. <i># Genes</i> indicates the number of genes from each list that were involved in the biological activity.</p