PBMC from RA and HD were stimulated in(A) or PMA/Iono (B). Subsequently, cytokine specific secretion assays (A) or intracellular stainings (B) were performed and cells were counterstained with anti-CD3, anti-CD4 and anti-CD8. Flow cytometric cytokine analysis were performed on live cells for IFNγ secretion (PI staining used for exclusion of dead cells) or on fixed cells for intra-cellular staining for IL-17, in each case after pre-gating on CD3+ T cells. A) Percentage of IFNγ producing T cells in samples from rheumatoid arthritis patients (n = 12, left panel and grey box in the right panel) and healthy donors (n = 13, clear box in the right panel) among CD4 SP, CD8 SP and CD4CD8 DP T cells. B) Percentage of IL-17 producing CD4 SP and CD4CD8 double positive T cells from RA patients (n = 4) and HD (n = 4). Presented box plots show median, interquartile range, and 10–90 percentiles. Significance as given, *p<0.05, **p<0.01, ***p<0.001.