Anti cd137 apc clone 17b5

TCR surface expression levels and activation marker expression of transduced 58αβ^−/− T cells were assessed by flow cytometry after incubating with anti‐TCRβ‐chain‐BV711 (clone H57‐597, Biolegend), anti‐CD69‐BV510 (clone H1‐2F3, Biolegend), and anti‐CD137‐APC (clone 17B5, Biolegend) diluted in PBS containing 2% FCS for 45 min at 4°C. 7AAD (Life technologies) was used to exclude dead cells. Flow cytometric analysis was performed according to the guidelines for the use of flow cytometry and cell sorting in immunological studies 23. Data acquisition was carried out with an AriaIII (BD Biosciences) and analyzed with FlowJo Version 10.4.1 (Tree Star).

To determine the expression of cell surface markers, cells were blocked with human Fc receptor (FcR) blocking reagent (Miltenyi Biotec) or Human TruStain FcX™ (BioLegend) and then stained with fluorochrome-conjugated antibodies diluted in flow cytometry staining buffer (PBS containing 2% FBS). Anti-CD137-PE (clone 4B4-1; BD Biosciences), anti-CD137L-APC (clone 5F4; BioLegend) or anti-CD137L-BV421 (clone 5F4; BioLegend), anti-CD45-PE/Dazzle CF594 (clone HI30; BioLegend) or anti-CD45-APC-H7 (clone 2D1; BD Biosciences), anti-CD25-BV421 (clone M-A251; BioLegend), anti-PD-1-APC (clone MIH4; eBioscience, Thermo Fisher Scientific) as where indicated. To determine the expression of mouse cell surface markers, cells were blocked with mouse FcR blocking reagent (Miltenyi Biotec) and stained with the following antibodies diluted in flow cytometry staining buffer: Anti-CD4-BUV395 (clone GK1.5, BD Biosciences) or anti-CD4-eFlour 450 (clone RM4-5, Thermo Fisher Scientific), anti-CD25-PE (clone PC61.5, Thermo Fisher Scientific) or anti-CD25-FITC (clone PC61.5.3, Thermo Fisher Scientific), anti-CD137-APC (clone 17B5; BioLegend), and anti-CD137L-PE (clone TKS-1; BioLegend). To determine cell viability, cells were stained with DAPI (Sigma-Aldrich) or LIVE/DEAD™ Fixable Violet Dead Cell Stain (Thermo Fisher Scientific) or LIVE/DEAD™ Fixable Near-IR Dead Cell Stain (Thermo Fisher Scientific).