< OMIPS Directory
OMIP-002

Phenotypic analysis of specific human CD8+ T-cells using peptide-MHC class I multimers for any of four epitopes

Pratip K Chattopadhyay, Mario Roederer, David A. Price Cytometry PART A, Volume 77A, Issue 9, 821-822 (2010)

PURPOSE: This panel was developed to determine the phenotype of human antigen (Ag)-specific CD8+ T-cells. Ag-specificities are identified by four peptide-major histocompatibility complex (MHC) class I (pMHCI) multimers (e.g., against Epstein-Barr virus (EBV) and cytomegalovirus (CMV) epitopes). Six markers of T-cell phenotype are used. This panel has been tested on fresh and cryopreserved peripheral blood mononuclear cells (PBMC), as well as bone marrow samples; staining may be performed in 96-well plates to increase throughput.

CELL TYPE: Fresh or cryopreserved PBMC, bone marrow mononuclear cells

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Violet viability
CD57 Human NK-1 FITC maturity
CD27 Human 1A4 PE-Cy5 maturity
CD127 Human R34.34 PE-Cy5.5 maturity
PD-1 Human MIH4 Biotin maturity
Biotin Human APC secondary reagent to PD-1
CD45RO Human UCHL1 APC-Alexa 700 maturity
CD3 Human SK7 APC-Cy7 T cell subset
CD14 Human M5E2 Pacific Blue exclusion
CD19 Human HIB19 Pacific Blue exclusion
CD4 Human Qdot 705 T cell subset
ccr7 Human 150503 PE-Alexa 750 maturity