OMIPs Directory

OMIP-042

21‐color flow cytometry to comprehensively immunophenotype major lymphocyte and myeloid subsets in human peripheral blood

Cytometry PART A, Volume 93, Issue 2, 186–189 (2018)
Karl W. Staser, William Eades, Jaebok Choi, Darja Karpova, John F. DiPersio

Description: This 21-color flow cytometry-based OMIP enables simultaneous quantification of monocytes, basophils, granulocytes, dendritic cells, natural killer cells, B cells, and all well-defined T and T helper cell subsets in the human peripheral blood. This panel captures the major phenotypes described in the NIH Human Immunology Project with additional markers for deep T cell analysis. We specifically designed this panel for analysis of peripheral blood from patients involved in our clinical trials of novel agents for the treatment of graft versus host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (alloHSCT). We have optimized this panel for the analysis of 1 ×10^6 fresh or previously frozen peripheral blood mononuclear cells (PBMCs).

Species: Human

Tissue: Peripheral blood

Cell Subset: B cell, T cell, T helper, T memory, Treg, Dc, Mdc/cdc, Pdc, Nk, Monocyte, Granulocyte, Basophil

Keyword: Allogeneic stem cell transplantation, Cryopreserved pbmc, Graft versus host disease (gvhd)

Machine: Bio-Rad Ze5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CCR10 Human 314305 APC Th subset T helper Low
Dead cells Human Live/Dead Fix Blue viability
CD16 Human 3G8 Brilliant Ultraviolet 496 monocyte monocyte, NK Med
CD56 Human NCAM16.2 Brilliant Ultraviolet 737 NK NK Low
CD20 Human L27 V450 B cell Med
CD4 Human SK3 Brilliant Violet 510 CD4 T cell T cell, T memory, T helper, Treg High
CD194 Human L291H4 Brilliant Violet 605 Th subset T cell, T helper Med
CD3 Human UCHT1 Alexa Fluor 488 T cell T cell, T memory, T helper, Treg High
CD183 Human 1C6 PE Th subset T cell, T helper Med
CD185 Human RF8B2 PE-Cy7 Th subset T cell, T helper Med
CD123 Human 32703 Alexa Fluor 700 pDC (plasmacytoid DC) DC Low
HLA DR Human G46-6 Brilliant Ultraviolet 661 DC DC High
CD8a Human RPA-T8 Brilliant Violet 650 CD8 T cell T cell High
CCR7 Human 150503 PE-CF594 central/effector T cell, T memory Low
CCR6 Human G034E3 Brilliant Violet 785 Th subset T cell, T helper Med
CD11c Human B-ly6 PE-Cy5 mDC DC Med
CD14 Human MfP9 Brilliant Ultraviolet 395 monocyte monocyte High
CD38 Human HIT2 Brilliant Violet 421 actvation T cell, T memory Low
CD25 Human 2A3 Brilliant Violet 711 Treg T cell, Treg Low
CD45RA Human HI100 PerCP-Cy5.5 naïve/memory T cell, T memory Med
CD127 Human RDR5 APC-eFluor 780 Treg T cell, Treg Med