Regulatory T Cells and Profile of FOXP3 Isoforms Expression in Peripheral Blood of Patients with Myelodysplastic Syndromes

Figure 1

Flow cytometry. The algorithm of regulatory T cells gating using PBMCs² of the 74-year-old healthy donor. (a) Lymphocytes among PBMCs. (b–c) Establishing FOXP3⁺ gate using CD3^- cells known not to express FOXP3: CD3^- cells among lymphocytes (b); establishing FOXP3 (exon 1)⁺ gate using CD3^- cells (c). (d–e) Detection of Treg: CD4⁺ T cells among lymphocytes (d); CD25⁺ FOXP3 (exon 1)⁺ – Treg among CD4⁺ T cells (e). (f–g) Analysis of FOXP3 isoform expression: all FOXP3⁺ cells among CD4⁺ T cells (f); calculation of FI (FOXP3 exon 2/total) parameter in all FOXP3⁺ cells = [(FOXP3 exon 2 MFI)/(FOXP3 exon 1 MFI)] (g). The solid arrows show the gating algorithm. The dashed bidirectional arrows show the alignment of the established gates for CD3^- and CD3⁺CD4⁺ cells. ¹a.u.: arbitrary units; ²PBMCs: peripheral blood mononuclear cells.