Immunophenotyping of human CD4+ and CD8+ T cell differentiation and exhaustion using flow cytometry (panel B)

MBTP 59 = Miltenyi Biotec-tested panel 59

This application protocol describes the analysis of human CD4+ and CD8+  T cell differentiation and exhaustion markers after T cell stimulation. To demonstrate this state-of-the-art panel for T cell differentiation as well as T cell exhaustion of both CD4 T cells and CD8 T cells, PBMCs were stimulated for 96 hours using Miltenyi Biotec’s T Cell TransAct™, human. The state of differentiation and exhaustion was defined by CD279 (PD-1), CD38, HLA-DR, and CD223 expression.

Protocol

Antibody panel for T cell differentiation and exhaustion

SpecificityClonePurposeFluorochromeDetection filter (nm) (laser)
7-AAD* –Viability –655–730 (blue)
CD279 (PD-1)REA1165T cell differentiation and exhaustionVio® Bright B515525/50 (blue)
CD4*REA623Helper T cellsVioGreen™525/50 (violet)
CD8*REA734Cytotoxic T cellsAPC-Vio 770750 LP (red)
CD223REA351T cell differentiation and exhaustionAPC655–730 (red)
CD3REA613T cellsVioBlue®450/50 (violet)
CD38REA572T cell differentiation and exhaustionPE585/40 (blue)
HLA-DRREA805T cell differentiation and exhaustionPE-Vio 770750 LP (blue)

* Alternative CE-IVD/ASR reagents available.

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Materials

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