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Comprehensive assessment of antigen-specific human T-cell functionality and memory

Andrew J. Graves, Marcelino G. Padilla, David A. Hokey Cytometry PART A, Volume 85, Issue 7, 576-579 (2014)

PURPOSE: This flow cytometry antibody panel was developed and optimized for the characterization of CD4+ and CD8+ T-cell memory and functional responses in adult and infant cryopreserved peripheral blood mononuclear cells (PBMC) stimulated with peptide pools to various antigens of interest. The panel has been used to evaluate Mycobacterium tuberculosis (TB) antigen-specific responses in clinical trial specimens, and is currently undergoing assay qualification.

CELL TYPE: Cryopreserved PBMC (adult and infant)


Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua viability
CD14 Human M5E2 V500 exclusion
CD19 Human HIB19 V500 exclusion
CD3 Human UCHT1 ECD T cell
CD4 Human RPA-T4 APC-eFluor 780 T cell subset
CD8 Human Hit8a Alexa Fluor 700 T cell subset
ccr7 Human G043H7 Brilliant Violet 605 Memory
CD45RO Human UCHL1 Brilliant Violet 785 Memory
IFN-gamma Human B27 V450 Th1
IL-2 Human MQ1-17H12 PE Th
TNF alpha Human Mab11 PE-Cy7 Th
IL-17A Human BL168 PerCP-Cy5.5 Th17
IL-22 Human IL22JOP APC Th22
CD107a Human H4A3 Alexa Fluor 488 Degranulation
CD40L Human TRAP1 PE-Cy5 Activation/B-Cell help