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Table 1.

The demographic and clinical characteristics of the participants.

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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+IFNγIL-17+, CD4+IFNγIL-22+, and CD4+IFNγIL-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.

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Figure 2.

The correlation between the percentages of Th17, Th22 and Th17/Th22 cells in patients.

The potential correlations among different subsets of CD4+ T cells were analyzed by Spearman rank correlation test. Data shown are the mean values of individual patients (n = 27). There was no significant correlation between the percentages of Th17 and Th17/Th22 cells in those patients.

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Figure 3.

The concentrations of plasma cytokines in the subjects.

The concentrations of plasma IL-17, IL-22, and IFNγ in individual subjects were measured by ELISA and the potential association of the concentrations of plasma cytokines with the percentages of corresponding cells was analyzed by Spearman rank correlation test. Data are expressed as the mean values of individual subjects (n = 27 for each group) from five separate experiments. A. ELISA analysis of the levels of plasma cytokines; B. Correlation analysis. The concentrations of plasma IL-17 were not correlated significantly with the percentages of Th17/Th22 cells in those patients (data not shown).

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Figure 4.

The concentrations of serum TSAb in the subjects.

The concentrations of serum TSAb in individual subjects were measured by ELISA, and the potential correlations between the levels of serum TSAb and the percentages of Th22, Th17 cells, and the levels of serum IL-17 and IL-22 in the GD patients were analyzed by Spearman rank correlation test. Data are expressed as the mean values of individual patients (n = 27). There was no significant association between the percentages of Th17, Th22, or the levels of plasma IL-17 and IL-22 and the levels of serum FT3, FT4, and TSH in those patients (data not shown).

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