Fig. 1 FACS analysis of the numbers of different subsets of circulating CD3+CD4+T cells in KD patients.
PBMCs were isolated from individual subjects and PBMCs 5*105/tube were stained in duplicate with PE-Cy5-anti-CD3 and PerCP-anti-CD4 or isotype controls, fixed and permeabilized, followed by intracellular staining with FITC-anti-IL-17, PE-Cy7-anti-IFN-γ, and PE-anti-IL-22. The frequency of CD3+CD4+IL-22+Th22, CD3+CD4+IL-17+Th17 and CD3+CD4+IFN-γ+Th1 cells were determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3+CD4+ cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3+CD4+IL-22+Th22, CD3+CD4+IL-17+Th17 and CD3+CD4+IFN-γ+Th1 cells were calculated, according to the total numbers of PBMCs, the frequency of different types of CD3+CD4+T cells. (A). Flow cytometry analysis; (B). The numbers of CD3+CD4+IL-22+Th22 cells; (C). CD3+CD4+IL-17+Th17 cells; (D). CD3+CD4+IFN-γ+Th1 cells. Data shown are representative FACS charts or the mean numbers of each type of cells per ml of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3+CD4+ T cells from individual groups of subjects (n=20 for the HC, n=17 for the patients with CAL, and n=26 for the patients with NCAL).