Table 1.
Demographics and clinical profiles of the CL patients and healthy volunteers in Morocco and Iran.
Fig 1.
Partial least squares-discriminant analysis (PLS-DA) modeling was applied for the classification of (a) Morocco CL patients infected with L. major and (b) Iran CL patients infected with L. tropica species compared to healthy volunteers.
A supervised global analysis of the 144 immune gene expression data was performed with the multivariate analysis tool PLS-DA. Each data point represents one sample and the circles denote respective group clusters.
Fig 2.
Volcano plot associated with differential immune gene expression in CL patients.
Volcano plots displaying significantly regulated genes in CL patients from (a) Morocco due to L. major and (b) Iran due to L. tropica compared to healthy volunteers. The y-axis corresponds to the P-values (-Log10), and the x-axis displays the Log2 fold change values. Using the P-value cutoff (P ≤ 0.05), the over- and under-expressed DEGs are depicted as red and blue dots, respectively. Black dots represent non-significantly different genes.
Fig 3.
Expression pattern of statistically significantly differentially expressed immune-related genes in CL patients compared to healthy volunteers.
(a) A heatmap displaying color-coded expression levels of genes in CL patients from Morocco (MA1-MA14) and Iran (IR1-IR16) infected with L. major and L. tropica, respectively, as well as healthy samples from each group (MAH1-MAH4; IRH1-IRH6). The heatmap was generated with normalized Log2 transformed dcRT-MLPA data. Only significant genes (10 and 19 DEGs were identified in Morocco and Iran studied groups, respectively) with a P-value cutoff (P ≤ 0.05) were included. It should be noted the CD14 and IFI6 are two common genes between both studied groups. The black color represents no detectable signal in the dcRT-MLPA assay. (b) The bar plot indicates Log2FC of DEGs associated with Morocco and Iran lesions. The red and blue colors of the bar plot represent relative significant increased or decreased expression of the genes in patients compared to healthy samples. The grey box colors indicate no significant changes in the expression of DEGs in the studied groups.
Fig 4.
Putative immune-related panels involved in skin lesion of CL patients infected with L. tropica and L. major.
DEGs obtained from (a) Moroccan patients infected with L. major and (b) Iranian patients infected with L. tropica relative to healthy volunteers. The X-axes indicate significant DEGs (P ≤ 0.05) and the Y-axes indicate the immune-related panels (specified in S1 Table). The bubbles’ sizes and colors correspond to Log2FC and the P-values of significant genes mapped to the indicated panels, respectively. Positive and negative Log2FC denote up- and down-regulated pathways, respectively. Larger bubble sizes refer to more significant genes.
Fig 5.
Similarities and differences in gene expression profile in CL patients infected with L. major and L. tropica species.
(a) A Venn diagram represents unique and common DEGs between Moroccan CL lesions infected with L. major and Iranian CL lesions infected with L. tropica compared to healthy samples. Genes expressed significantly higher or lower than the control are shown in red or blue fonts, respectively. (b) A heatmap displaying color-coded expression levels of genes in CL patients from Morocco (MA1-MA14) and Iran (IR1-IR16) infected with L. major and L. tropica, respectively, as well as healthy samples from each nation (MAH1-MAH4; IRH1-IRH6). The heatmap was generated with normalized Log2 transformed dcRT-MLPA data. The gene expression pattern of two common genes (CD14 and IFI6) expression in L. major and L. tropica-infected patients compared to healthy volunteers, as shown by a heatmap on the left and a bar chart on the right.