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Validated multifunctional characterization of antigen-specific human T cells by intracellular cytokine staining

Stephen C. De Rosa, Donald K. Carter, M. Juliana McElrath Cytometry PART A, Volume 81A, Issue 5, 362-363 (2012)

PURPOSE: This panel was developed, optimized, and validated for assessment of CD4+ and CD8+ T-cell responses to various peptide pools for antigens of interest in cryopreserved peripheral blood mononuclear cells (PBMC) from adult humans (Table 1). The panel has been used to evaluate HIV- and TB-specific responses to candidate vaccines for these pathogens, although the panel can be used with peptide pools for any proteins. The panel has not been tested with freshly-isolated PBMC or with whole blood.

CELL TYPE: Cryopreserved PBMC


Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua viability
CD14 Human Tük4 Qdot 655 exclusion
IFN-gamma Human B27 V450 function
IL-2 Human MQ1-17H12 PE function
TNF alpha Human Mab11 FITC function
IL-4 Human MP4-25D2 APC function
MIP-1 beta Human D21-1351 Alexa Fluor 700 function
CD40L Human TRAP1 PE-Cy5 function
CD107a Human H4A3 PE-Cy7 function
CD3 Human UCHT1 PE-Texas Red lineage
CD4 Human 13B8.2 APC-Alexa 750 lineage
CD8 Human SK1 PerCP-Cy5.5 lineage