Figure 1. Flow cytometry analysis of different subsets of CD4+ T cells.

PBMCs were isolated form individual participants and stimulated with, or without, PMA/ionomycin and harvested. The cells were stained with APC-anti-CD4, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17, PE-Cy7-anti-IFNγ, and PE-anti-IL-22 and flow cytometry. Subsequently, the cells were gated first on CD4⁺ cells for analysis of the frequency of CD4⁺IFNγ⁺ and CD4⁺IFNγ⁻ cells. The CD4⁺IFNγ⁺ cells were further analyzed for CD4⁺IFNγ⁺IL-17⁺ cells (column I), while the CD4⁺IFNγ⁻ cells were further analyzed for CD4⁺IFNγ⁻IL-17⁺, CD4⁺IFNγ⁻IL-22⁺, and CD4⁺IFNγ⁻IL-17⁺IL-22⁺ cells (column II), followed by quantitative analyses. Data are representative charts or expressed as the mean values of individual participants from sequential experiments. A. Representative charts of flow cytometry analysis; B. Quantitative analysis.