Figure 2. The percentages of circulating Th22, total Th17, and IL-22⁺Th17, IL22⁺Th1, but not total Th1 cells decreased following MTX+LEF therapy only in patients that exhibited clinical improvement (n = 40).

Peripheral blood mononuclear cells (PBMCs) were collected from RA patients responsive to treatment at baseline (before treatment, 0 week) and after treatment (12 weeks) with MTX+LEF, and analyzed by flow cytometry for the percentage of different Th subsets, including IFN-γ⁻IL-17⁻IL-22⁺ (Th22), IFN-γ⁻IL-17⁺ (total Th17), IFN-γ⁻IL-17⁺IL-22⁺ (IL-22⁺Th17), IFN-γ⁺IL-17⁻ (total Th1), and IFN-γ⁺IL-17⁻IL-22⁺ (IL-22⁺Th1) cells. (A) The gating strategy for detecting different subsets of Th cells and representative flow images on samples from RA patients at 0 week (0 W), 12 week (12 W), or from healthy controls (HC; n = 20). R1, lymphoctyes; R2, IFN-γ⁻CD4⁺T cells; R3, IL-17⁻CD4⁺T cells; I, Th22 cells; II, IL-22⁺Th17 cells; III, IL-22⁻Th17 cells; IV; IL-22⁺Th1 cells; V; IL-22⁻Th1 cells.