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OMIP-066

Identification of Novel Subpopulations of Human Group 2 Innate Lymphoid Cells in Peripheral Blood

Yoichiro Ohne Cytometry PART A, Volume 97, Issue 10, 1028-1031 (2020)

PURPOSE: This 14‐color flow cytometry panel was designed to identify newly described subpopulations within human group 2 innate lymphoid cells (ILC2s) and other ILC subsets. This panel also allowed to identify recently reported subpopulations of peripheral blood CRTH2− c‐Kit+ ILCs. We validated this panel mostly in human peripheral blood but also confirmed that the same panel and gating strategy works well in human tonsillar cells. The panel contains a few markers indicating the activation status of ILCs. In addition, phycoerythrin (PE) channel is available for the markers of interest in each study. In the validation studies described here, PE channel was used to test the expression of some markers. These features make this panel applicable for immunophenotyping of ILCs in various disease states. © 2020 International Society for Advancement of Cytometry

CELL TYPE: PBMC

MACHINE: BD FACSymphony A5

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Blue Viability
CCR10 Human 1B5 BUV395 Subpopulation
CD45 Human HI30 BUV805 Lymphocyte
CD3 Human SK7 Brilliant Violet 510 Lineage/Dump
CD4 Human RPA-T4 Brilliant Violet 510 Lineage/Dump
CD14 Human M5E2 Brilliant Violet 510 Lineage/Dump
CD19 Human Brilliant Violet 510 Lineage/Dump
CD34 Human 581 Brilliant Violet 510 Lineage/Dump
CD123 Human 6H6 Brilliant Violet 510 Lineage/Dump
CCR6 Human G034E3 Brilliant Violet 421 Lineage/Dump
KLRG1 Human 13F12F2 SuperBright 702 Subpopulation
CD336 Human p44-8 Brilliant Violet 786 ILC3 in tonsil
CD127 Human REA614 VioBright 515 Total ILC
TSLP Receptor Human 1F11 BB700 ILC2 activation
NKp46 Human DX22 PE-Dazzle 594 Subpopulation
CD117 Human 104D2 PE-Cy7 Subpopulation
CD294 Human BM16 Alexa Fluor 647 ILC2
CD56 Human NCAM16.2 APC-R700 Subpopulation
CD94 Human APC-Fire 750 NK