FIGURE 1. T2D patients have elevated percentages of circulating Th17 cells.

A, One representative flow plot of 10 ND and 11 T2D whole blood samples stained for CD3⁺ CCR4⁺ CCR6⁺ cells. Blood donors for these studies are described in Table II. T cells were identified based on forward and side scatter and CD3 staining (not shown) and plotted to show CCR4 (x-axis) and CCR6 (y-axis) staining. Note that slight elevation of CCR6 expression in lower left quadrant was inconsistent. Percent double-positive cells are indicated in the top right quadrant. B, Summary of flow cytometric analyses of percentage CCR4/CCR6 double-positive T cells in whole blood. Each dot represents one individual. n = 10 for ND or 11 for T2D samples. C, Representative flow plot of T cell purity for fresh ex vivo T cells used in purified T cell analyses. T cells in this sample are 98.8% pure based on CD3 expression (top left quadrant). CD69 is shown on the x-axis, and CD3 is displayed on y-axis, the former to indicate low T cell activation levels. D, ChIP measuring constitutive association of acetylated histone H3 with the IL-17 promoter in fresh ex vivo T cells from ND (white bar) or T2D (black bar) donors. Bars show average signal (± SEM) compared with control immunoprecipitations with anti-GST Ab. p values calculated by Mann–Whitney U test highlight the significant difference between ND and T2D samples. n = 4 to 5 independent determinations. E, Relationship between BMI (x-axis) and percentage of CD3⁺ CCR4⁺ CCR6⁺ cells (y-axis) in ND (left panel) or T2D (right panel) cohorts. Correlation was calculated by a Pearson’s test. Pearson’s r value is shown below the significant (<0.05) p value.