Fig. 8

Human macrophages treated with hypoxic GBM-derived EVs have lower capacity to induce T cell activation markers after superantigen stimulation. A., B. Human MDMs from two donors (Donor 1 and Donor 2) were cultured for 7 days in the absence (M) or presence (MEV) of hypoxic GBM-derived EVs (1% EVs). The macrophages were then washed and co-cultured with CD3⁺ T cells from the same donor for 3 days with or without the addition of Staphylococcal Enterotoxin B (1 μg/ml, SEB 1 or 0.1 μg/ml, SEB 0.1). On day 1 and day 3 cells were harvested and the expression of (A) CD69 and (B) CD25 on T cells (CD4⁺ and CD8⁺) was measured by flow cytometry. Values are expressed as mean ± SD of three replicates for each donor, and comparisons were made using an unpaired t test. *p < 0.05, **p < 0.005, ***P < 0.001