Fig. 1

Frequencies of NK and NKT cells; CD4⁺, CD8⁺, double positive (DP) and double negative (DN) CD3⁺ T-lymphocyte subpopulations. a-f Flow cytometry-representative protocol: Cells obtained from cutaneous leishmaniasis lesions were stained with anti-CD3, anti-CD4, anti-CD8, anti-CD56, anti-CD107a and 7-AAD. Lymphocytes region was created on forward (FSC) vs side (SSC) scatter dot-plot (a). Cells gated on [Lymphs.1] were analyzed through dot-plots SSC-Height vs SSC-Area (b) and SSC-Height vs SSC-Width (c) to exclude doublets and debris. d Dead cells (7-AAD⁺) were excluded from the analysis and a gate encompassing 7-AAD^neg cells was performed. e Based on this gate, NK cells (CD56⁺CD3^neg), NKT (CD56⁺CD3⁺) and T lymphocytes (CD56^negCD3⁺) were identified. f Based on T lymphocyte gate (T Lymph), CD4, CD8, double-negative (DN) and double-positive (DP) T lymphocytes were determined. g Bar graphs representing the mean ± SEM of percentages of NKT and NK cells; CD4⁺, DP, CD8⁺ and DN T lymphocytes obtained from cutaneous leishmaniasis lesions of 18 patients. Statistical analyses were performed using ANOVA test and Dunn’s post-hoc test. Results were considered significant with P < 0.05 (*P < 0.05; **P < 0.01; ***P < 0.001)