Fig. 3: T cell activation enhances TF-mediated CD4⁺ T cell thrombogenicity.

CD4⁺ T cells were isolated from donor human blood and plated at a density of 0.8 × 10⁶/ml with IL-2 for unactivated conditions (θ). Plated cells were activated by anti-CD3/anti-CD28 activation beads and stimulated with IL-2 for Th0 conditions, and differentiation cytokines (αIL-4 + IL-12) were added to skew cells to a Th1 lineage. a–d Following 5 days in culture, θ, Th0 and Th1 cells were washed with EDTA-containing PBS, and their ability to initiate thrombin generation was analysed by calibrated automated thrombinography in normal pooled platelet-poor plasma. b Lagtime, c peak thrombin levels and d endogenous thrombin potential (ETP) were measured and compared between θ, Th0 and Th1 cells. e The rate of clot formation was measured in θ, Th0 and Th1 cells. f–i θ, Th0 and Th1 cell-mediated thrombin generation was analysed by calibrated automated thrombinography using FVII-deficient platelet-poor plasma. g Lagtime, h peak thrombin levels and i ETP were measured and compared between θ, Th0 and Th1 cells. j F3 gene expression, k, l the percentage of cells expressing TF, m cell surface TF expression and n T cell-dependent FXa generation was measured in θ, Th0 and Th1 cells. Student’s paired t-test (two-tailed) (c–e, g–i, l–n), Wilcoxon test (two-tailed) (b), or Mann–Whitney U Test (two-tailed) (j) was used to determine statistical significance. Data is expressed as mean ± s.d. (b–e, g–i, l–n) for 7 (a–d), 10 (e), 6 (g–i), 8 (j) and 4 (l–n) biological donors/group. Source data are provided in the Source Data file.