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. 2014 Aug 1;124(9):4004–4016. doi: 10.1172/JCI75051

Figure 1. EMRA CD8+ T cells exhibit characteristics of senescent T cells.

Figure 1

(A) Multiparameter flow cytometry examining expression of the senescence features KLRG1, CD57, and γH2AX in the 4 CD45RA/CD27-defined CD8+ T cell subsets after an 8-hour stimulation with 0.5 μg/ml anti-CD3. Pie charts show the average of 7 donors. N, naive. (B) Representative blot showing telomerase activity on day 3 after activation with 0.5 μg/ml anti-CD3 and 5 ng/ml IL-2 (n = 4). Lanes were run on the same gel but were noncontiguous (black line). Telomerase activity was calculated by densitometry and expressed in arbitrary units relative to the internal standard (IS). (C) Representative immunoblots of p-p38 and total p38 together with β-actin after a 30-minute stimulation with 0.5 μg/ml anti-CD3 and 5 ng/ml IL-2. (D) Phosphoflow data after a 30-minute stimulation with 0.5 μg/ml PMA and ionomycin. Data are shown relative to the naive subset, n = 9. (E and F) ROS production, obtained using MitoSox (E) and DHE (F), in the CD8+ T cell subsets after overnight stimulation (n = 8). (G) Proliferation of CD8+ T cell subsets, assessed by Ki67 staining, after stimulation with 0.5 μg/ml anti-CD3 and 5 ng/ml IL-2 for 3 days. Horizontal lines depict means. (H) Multiparameter flow cytometry examining expression of IFN-γ, TNF-α, perforin, and granzyme B after an 8-hour stimulation with 0.5 μg/ml anti-CD3. Pie charts show the average of 7 donors. *P < 0.05, **P < 0.01, ***P < 0.001, repeated-measures ANOVA followed by Tukey correction.