Figure 1. Flow cytometric analysis of T cell subsets.

PBMCs were stained with anti-CD4 PE-cy7, anti-CD25 APC, anti-IFN-γ FITC, anti-IL-17 PE, anti-IL-4 APC and anti-Foxp3 FITC. CD4+ cells were gated for further analysis. PBMC from patients before KT, patients at 1month after KT and patients at 3 month after KT were stimulated for 4 h ex vivo with PMA and ionomycin in the presence of Golgi Stop. The percentage of Target cells was measured by flowcytometry. The frequency (%) of Lymphocyte/Leukocyte cells, CD4⁺T/Lymphocyte cells, IL-17+/CD4⁺T cells, IFN-γ⁺/CD4⁺T cells, IL-4⁺/CD4⁺T cells (A) and CD25⁺FOXP3⁺/CD4⁺T cells (B) in patients before KT, patients at 1month after KT and patients at 3 month after KT. After surface staining with anti-CD4, CD45 and CCR7 mAbs, cells were fixated and permeabilized and intracellular accumulated cytokines were detected with IL-17 mAbs. T_naïve/CD4⁺T (CD45RA⁺CCR7⁺/CD4⁺Tcells), IL-17⁺/T_naïve, T_CM/CD4⁺ T (CD45RA⁻CCR7⁺/CD4⁺Tcells), IL-17⁺/T_CM ⁺ and T_EM/CD4⁺T (CD45RA⁻CCR7⁻/CD4⁺Tcells), IL-17⁺/T_EM ⁺ (C).