Immunophenotyping of human CD4+ and CD8+ T cell activation and exhaustion using flow cytometry

MBTP 55  = Miltenyi Biotec-tested panel 55

This application protocol describes the analysis of human CD4+ and CD8+ T cell activation and exhaustion markers after T cell stimulation. To demonstrate this state-of-the-art panel for T cell activation as well as T cell exhaustion of both CD4 T cells and CD8 T cells, PBMCs were stimulated using Miltenyi Biotec’s T Cell TransAct™, human for 72 hours.  The state of activation and exhaustion was defined by CD25, CD223, CD279 (PD-1), and CD69 expression.

Protocol

Antibody panel for T cell activation and exhaustion

SpecificityClonePurposeFluorochromeDetection filter (nm) (laser)
7-AAD*-Viability-655−730 (blue)
CD279 (PD-1)REA1165T cell activation and exhaustionVio® Bright B515525/50 (blue)
CD4*REA623Helper T cellsVioGreen™525/50 (violet)
CD8*REA734Cytotoxic T cellsAPC-Vio 770750 LP (red)
CD223REA351T cell activation and exhaustionAPC655−730 (red)
CD3REA613T cellsVioBlue®450/50 (violet)
CD25REA570T cell activation and exhaustionPE585/40 (blue)
CD69REA824T cell activation and exhaustionPE-Vio 770750 LP (blue) 

* Alternative CE-IVD/ASR reagents available. 

Gating strategy

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Materials

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