ADAM17 Inhibition in Activated CD3+ T Cells

CD3⁺ T cells were isolated from PBMCs using a MojoSort^TM Human CD3 T Cell Isolation Kit (BioLegend, USA). The purity of isolated CD3⁺ T cells was above 95%, as examined by flow cytometry. ADAM17 inhibition experiment was carried out according to previous report [41 (link)]. Isolated CD3⁺ T cells were cultured in X–VIVOTM 15 medium containing 20% FBS, and treated for 1 h in the presence or absence of TAPI-1 (20 mM). Next, they were incubated on a plate coated with anti-CD3 antibody (10 μg/ml) and anti-CD28 antibody (1 μg/ml) for 4 h. Cells were harvested after treatment, and flow cytometry was performed to measure the fluorescence intensity of ADAM17 and IL-23 R in CD4⁺ T cells.