Immunophenotyping of human tumor-reactive T cell functionality using flow cytometry

MBTP 61 = Miltenyi Biotec-tested panel 61

This application protocol describes the analysis of human tumor-reactive T cell functionality by detection of intracellular and extracellular markers. To demonstrate the state-of-the-art panel, tumor-infiltrating leukocytes (TILs) were isolated from a human melanoma digest. The contained tumor-reactive T cells (TRTs) were enriched using CD137 MicroBeads and expanded for two weeks using the CliniMACS Prodigy®. For functionality testing, TRTs were then co-cultured with autologous or mismatched tumor cell lines in presence of a protein transport inhibitor containing monensin and brefeldin A. TRTs were subsequently analyzed by flow cytometry to determine their anti-tumor reactivity in terms of T cell degranulation (CD107a/LAMP-1), T cell activation (CD154, CD137), and production of inflammatory cytokines (IFNγ, TNFα). 


Antibody panel for human tumor-reactive T cell functionality

SpecificityClonePurposeFluorochromeDetection filter (nm) (laser)
CD4REA623Helper T cellsFITC525/50 (blue)
CD8REA734Cytotoxic T cellsVioGreen™525/50 (violet)
IFNγ REA600Inflammatory cytokineAPC-Vio® 770750 LP (red)
CD154REA238T cell activationAPC655–730 (red)
CD107a/LAMP-1REA792T cell degranulationVioBlue®450/50 (violet)
CD137REA765T cell activationPE585/40 (blue)
TNFαcA2Inflammatory cytokinePE-Vio 770750 LP (blue) 

Gating strategy

Matching products:


Matching products:

Seems like you are coming from USA!
Do you want to visit our website in your country?