Flow cytometry gating strategy and frequency of TcTLE-specific CD8⁺ T cells.

<p>Dot plots representative of the analysis of an HLA-A2-negative chagasic patient. (A) Lymphocytes were selected, and cell doublets were excluded from the analysis for the identification of CD8⁺ T cells based on the forward scatter area (FSC-A) and forward scatter height (FSC-H) (singlet 1), forward scatter width (FSC-W) and forward scatter height (FSC-H) (singlet 2), and side scatter width (SSC-W) and SSC-H characteristics (singlet 3). Finally, live CD3⁺ CD8⁺ cells were selected. (B) Selection of TcTLE peptide-specific CD8⁺ T cells by soluble tetramer assay. (C) Frequency of TcTLE peptide-specific CD8⁺ T cells from 20 HLA-A2-negative chronic chagasic patients. The cut-off point, which was fixed at 0.063, is indicated by the dotted line.