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. 2022 Sep 12;13:827786. doi: 10.3389/fimmu.2022.827786

Figure 1.

Figure 1

Activated EC induce CD69 expression in T cells, without proliferation or activation. HMEC were left unstimulated or stimulated with TNFα and IFNγ for 3 days before addition of FACS-sorted CD3+ T cells to the co-culture, in the presence or absence of soluble anti-CD3 stimulation. Conventional dendritic cells (cDC) were added as a positive control to induce T cell proliferation. CD69 expression was analyzed by flow cytometry after 4 days of co-culture. (A) Representative flow cytometry plot, (B) percentage of positive cells and median fluorescent intensity (MFI). Kruskal-Wallis with Dunn’s post-hoc test compared to unstimulated, c.q. only anti-CD3 stimulated T cells. *p < 0.05. (C) Proliferation was assessed by CellTrace Violet (CTV) dilution assay. Kruskal-Wallis with Dunn’s post-hoc test. (D) Expression of CD25 and Ki67 after 4 days of co-culture with unstimulated T cells, resting/activated HMEC or anti-CD3/CD28 beads. N = 3, mean+SEM. Kruskal-Wallis with Dunn’s post-hoc test. *p < 0.05, **p < 0.01.