The expansion of EBV-specific T cells and their detection were described elsewhere (Izawa et al., 2017 (link)). In brief, PBMCs were co-cultured with 40-Gy irradiated autologous or HLA-A matched LCLs at a 40:1 ratio (PBMCs: LCLs). Cells were re-stimulated with irradiated LCLs at a ratio of 4:1 on days 9, 16, and 23. IL-2 (50 U/ml) was added from day 16 of culture. For the detection of EBV-specific CD8+ T cells, cells were incubated with unlabeled Pro5 MHC Pentamer mix followed by Pro5 fluorotag APC, anti–CD3-BV785 (Proimmune), anti–CD4-BV421, anti–CD8-BV510, anti–CD19-BV711 (from BioLengend), and anti–CD137-PE (from BD Pharmingen). Pentamer mix contains HLA A*02:01/YLLEMLWRL (residues 125–133 from LMP1), HLA A*02:01/FLYALALLL (residues 356–364 from LMP2), HLA A*02:01/CLGGLLTMV (residues 259–267 from LMP2), and HLA A*02:01/GLCTLVAML (residues 259–267 from BMLF-1). Stained cells were analyzed by flow cytometry. Three LCLs from DiGeorges patients were retrieved from our hospital and other places. Only one was HLA-A02*.
Anti cd137 pe
Manufactured by BD
Anti-CD137-PE is a fluorescently-labeled antibody that binds to the CD137 receptor, also known as 4-1BB, on the surface of cells. It can be used for the detection and analysis of CD137-expressing cells in flow cytometry applications.
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2 protocols using anti cd137 pe
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EBV-specific T cell expansion and detection
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Multiparametric Immune Cell Profiling
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Epitope-specific cell lines were stained with anti-CD3, anti-CD4, anti-CD8, anti-CD45RO and anti-CCR7 for 15 min at 4 °C. Anti-CD107a-BV421 (328,626, BioLegend) was added during culture. For intracellular staining, cells were fixed, permeabilized, and stained with anti-CD137-PE (555,956, BD), anti-IFN-γ and anti-TNF using the Foxp3/transcription factor staining buffer set. Cells were acquired on an LSRFortessaX20.
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