Figure 1.

FACS analysis of the numbers of different subsets of circulating CD3⁺CD4⁺T cells and ELISA analysis of serum IL-10 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10⁵/tube were stained in duplicate with FITC-anti-CD3, PE-Cy7-anti-CD25, and PerCP-anti-CD4 or isotype controls, fixed, and permeabilized, followed by intracellular staining with PE-anti-Foxp3. The frequency of CD3⁺CD4⁺CD25⁻Foxp3⁺ and CD3⁺CD4⁺ CD25⁺Foxp3⁺T cells was 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⁺Foxp3⁺T cells were calculated, according to the total numbers of PBMCs and the frequency of different types of CD3⁺CD4⁺Foxp3⁺T cells. The concentrations of serum IL-10 in individual subjects were determined by ELISA. (a) Flow cytometry analysis; (b) the numbers of CD3⁺CD4⁺CD25⁺Foxp3⁺T cells; (c) the numbers of CD3⁺CD⁴⁺CD25⁻Foxp3⁺T cells; (d) serum levels of IL-10. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood and the mean levels of serum IL-10 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 and serum IL-10 from individual groups of subjects (n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).