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Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV+
human peripheral blood mononuclear cells (PBMCs) were restimulated for 4 hours either with 20 μL/mL of reconstituted PepTivator®
BKV LT1 (A), 20 µL/mL BKV VP1 (B), or a mixture of both peptide pools (C). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4+
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