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Pan-leukocyte immunophenotypic characterization of PBMC subsets in human samples

Gemma Moncunill, Hannah Han, Carlota Dobano, M. Juliana McElrath, Stephen C. De Rosa Cytometry PART A, Volume 85, Issue 12, 995-998 (2014)

PURPOSE: This phenotyping panel was developed to measure the relative frequencies of multiple leukocyte cell subsets in peripheral blood mononuclear cells (PBMC) from African infants and children, including the expression of immune activation and differentiation markers. It was optimized with the objective of obtaining the maximum information concerning the immune status and cell subsets that could influence the immune response to vaccines and infectious diseases using small volumes of pediatric samples. It was developed using cryopreserved PBMC from healthy HIV-uninfected and HIV-infected US adults, but it has also been tested with cryopreserved PBMC from US infants. Although we have not tested the panel on whole blood, it is likely that the panel could be used with whole blood following minimal optimization.

CELL TYPE: Cryopreserved PBMC from adults and infants


Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua viability
CD4 Human SK3 BUV395 T cell
CD19 Human SJ25C1 BUV737 B cell
CD25 Human M-A251 Brilliant Violet 421 Treg
HLA-DR Human Brilliant Violet 570 Activation
CD56 Human HCD56 Brilliant Violet 605 NK, NKT-like cell
CD45RA Human HI100 Brilliant Violet 650 memory,differentiation
CD14 Human MfP9 Brilliant Violet 711 monocyte
ccr7 Human G043H7 Brilliant Violet 785 n/a
CD57 Human NK-1 FITC n/a
CD8 Human SK1 PerCP-Cy5.5 T cell lineage
TCR V delta 2 Human B6 PE ?d T cell
CD3 Human UCHT1 ECD T cell lineage
CD38 Human HIT2 PE-Cy5 Activation, plasmablasts
TCR gamma/delta Human 11F2 PE-Cy7 ?d T cell
CD127 Human A019D5 APC Treg, memory, differentiation
CD159c Human 134591 Alexa Fluor 700 NK receptor
CD16 Human 3G8 APC-Cy7 NK, monocyte