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. 2018 Mar 12;9:454. doi: 10.3389/fimmu.2018.00454

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Copyright © 2018 Pinto, Medeiros, Teixeira-Carvalho, Eloi-Santos, Fontes-Cal, Rocha, Dutra, Correa-Oliveira and Gomes.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of lymphocytes and subsets. Flow cytometry gate strategy of non-infected (NI) individuals is represented. Frequency of CD4⁺ lymphocytes expressing CD25, CD28, and CTLA-4 (A) in culture without stimulation (control) and after in vitro stimulation with T. cruzi antigens (TRIPO). Frequency of Th1, Th2, Th17, and T regulatory (Treg) lymphocytes subsets (B) and, proportion of lymphocyte subsets (C). The groups evaluated were NI individuals (n = 6), indeterminate (IND, n = 10), and cardiac (CARD, n = 9) clinical forms of Chagas disease. Significant differences (p < 0.05) between groups are evidenced by lines and asterisks (*) according Kruskal–Wallis test, followed by Dunn’s post hoc test. The # sign indicates the significant difference between control and TRIPO cultures using Wilcoxon signed-rank test. Boxes show the median and interquartile ranges, whiskers indicate the highest and lowest observation, and dots represent the outliers.