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. 2022 Jul 18;9:865330. doi: 10.3389/fmed.2022.865330

Figure 1.

Figure 1

Gating strategies and frequency evaluation of regulatory T cells subsets. To exclude cell doublets and debris from data, cells were gated on singlet regions through dot plot FSC-H vs FSC-A (A); Next analysis was initiated from singlets and gated in live and dead histogram, for viability evaluation (B); lymphocyte population were determined from viable cells in SSC-A vs FSC-A dot-plot (C); the dot-plot from lymphocyte cells was used for CD4+ T cell determination (D); CD25 vs Foxp3 gated on CD4+ was used to define CD4+ T reg cells (E). Graphical representation of CD4+CD25+Foxp3+ frequencies in different clinical forms of leprosy (F). From lymphocyte dot-plot, CD8+ cells were gated (G) and CD25 vs Foxp3 were defined (H) graphical representing of CD8+CD25+Foxp3+ cells (I); Ratio determination of CD4+CD25+Foxp3/CD8+CD25+Foxp3+ frequencies in PBMC (J). In (F,I) each symbol represents an individual studied according to the group; horizontal lines represent the mean. In (J) each bar represents the mean and standard deviation of the groups and the culture condition. One-way Anova was used for statistical analysis where, *p ≤ 0.05, **p leq 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001. T1R, Type 1 Reaction; BL, borderline lepromatous; T2R, Type 2 Reaction; LL, polar lepromatous patients; HV, healthy volunteers.