< OMIPS Directory
OMIP-021

Simultaneous quantification of human conventional and innate-like T-cell subsets

Nicholas A. Gherardin, David S. Ritchie, Dale I. Godfrey, Paul J. Neeson Cytometry PART A, Volume 85, Issue 7, 573-575 (2014)

PURPOSE: This panel was developed in order to simultaneously quantify both conventional peptide-MHC-restricted, and innate-like T-cell compartments in human peripheral blood samples. The panel can assess the dynamics of naïve through to terminally differentiated effector memory T-cell subsets, as well as enumerating natural killer T (NKT) cells, mucosal-associated invariant T (MAIT) cells, γδ T-cells, and subsets thereof. The panel is suitable for use on both, freshly isolated or cryopreserved peripheral blood mononuclear cells (PBMC). Staining may be performed in a 96-well plate to increase throughput.

CELL TYPE: PBMC

MACHINE: BD LSR Fortessa

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua Viability (dump)
CD3 Human OKT3 Brilliant Violet 785 Lineage
CD4 Human RPA-T4 APC-Cy7 Phenotyping
CD8a Human RPA-T8 Brilliant Violet 650 Phenotyping
CD8b Human 2ST8.5H7 APC Phenotyping
CD27 Human O323 Brilliant Violet 711 memory T cell subset
CD28 Human 28.2 PE-Cy5 memory T cell subset
CD45RA Human HI100 PerCP-Cy5.5 memory T cell subset
CD45RO Human UCHL1 Alexa Fluor 700 memory T cell subset
CD161 Human HP-3G10 Brilliant Violet 605 MAIT
ccr7 Human 3D12 PE-Cy7 memory T cell subset
TCR gamma/delta Human 11F2 FITC ?d T cell
TCR V alpha 7.2 Human 3C10 PE MAIT
Streptavidin Human Brilliant Violet 421 NKT