Fig. 2

Changes in peripheral blood regulatory T-cells (Treg) frequencies in response to 20 weeks of exercise intervention in older adults with RA. a. CD4+ cells were gated from the total number of lymphocytes and then b. further gated into CD127-CD25+. The Foxp3+ Tregs were identified within the CD127-CD25+ and the gate was set based on lack of expression in the CD4 + CD25- cells. c. effector T cells were identified by a CD25+ gate, then a Foxp3- gate set adjacent to the Foxp3+ gate. Paired measurements of d. CD127-CD25 + Foxp3+ Tregs, e. CD25 + Foxp3- effector T cells frequencies in blood and f. absolute number of Tregs/ml, pre and post 20 weeks of control (n = 24) or exercise (n = 24) intervention. Frequencies are related to the CD4+ gate. g. Flow cytometry gating of proliferating CD4+ T cells after 4 days of stimulation with anti-CD3/anti-CD28 and h. CellTrace violet staining of CD4+ T cells from matched samples collected pre (n = 4) and post exercise (n = 4) with i. graphs showing % dividing cells and proliferation index analysed by the FlowJo proliferation analysis tool and j. cytokines in cell culture supernatants. P values were determined using Wilcoxon signed-rank test comparing data from samples collected at baseline (pre) and after 20 weeks (post). The Mann-Whitney U test was used for comparison between the two groups. A ratio paired t-test was used to compare data from the T cell proliferation assay between samples collected at baseline (pre) and after 20 weeks (post). *p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001 **** p ≤ 0.0001