Chiocca Lab

Downloadable Figure: GBM EVs block T-cell receptor mediated T-cell activation.

A. FACS-gating strategy. Cells were gated by FSC/SCC while excluding duplets by FSC-A/FSC-H. Subsequently, CD3+CD56- T cells were gated with further classification of CD4+ and CD8+ T cells. Finally CD4+ T cells and CD8+ T cells were measured for their CD69, CD25, PD1 and TIM3 expression. B. PD-L1^High and PD-L1^low GSC EVs do not activate T-cells measured by CD69 and CD25 expression. PD-L1^High and PD-L1^low GSC EVs inhibit CD4+ T-cell % CD69 and CD25 and CD69⁺CD25⁺ expression change compared to anti-CD3 for 2d ± GBM EVs (5µg/ml). OKT3-stimulated PBMCs were from 8 human (P1-8) volunteers ± PDL1^high/low EVs, NSC-EVs were added to OKT3-stimulated PBMCs from 3 human volunteers). D. Examples of CD3+CD8+ T-cell CD69 and CD25 expression changes due to PD-L1^High and PD-L1^low treatments measured via FACS. E. PD-L1^High EVs suppress T-cell activation in a PD1 dependent matter. Examples of CD3+CD4+ and CD3+CD8+ T-cell CD69 and CD25 expression changes due to PD-L1^High and PD-L1^low treatments and PD1-Blockade are shown.