OMIPs Directory

OMIP-064

A 27‐Color Flow Cytometry Panel to Detect and Characterize Human NK Cells and Other Innate Lymphoid Cell Subsets, MAIT Cells, and γδ T Cells

Cytometry PART A, Volume 97, Issue 10, 1019-1023 (2020)
Nina Hertoghs, Katharine V. Schwedhelm, Kenneth D. Stuart, Margaret Juliana McElrath, Stephen C. De Rosa

Description: This 27‐color flow cytometry panel was developed in order to assess immunological changes over the course of an immunization and challenge regimen in two experimental malaria vaccine trials. The aim of the study was to find correlates of vaccine‐induced protection. Several studies have indicated that protection against malaria appears to involve immune responses at various immunological sites, with liver‐resident responses playing an essential role. As it is not feasible to monitor the immune responses within the liver in humans, this panel is developed with the aim to thoroughly characterize the immune responses over time in blood in addition to detecting changes that might reflect what happens in other immunological sites like the liver. The focus of this panel is to detect several innate lymphoid cell populations, including NK cells and their activation status. Moreover, unconventional T cells like mucosal associated invariant T cells and γδ T cells are assessed in the panel.

Species: Human

Tissue: Peripheral blood

Cell Subset: Mait, Tcrgd, Ilc, Nk, Monocyte

Keyword: Activation, Malaria, Vaccination

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CD159c Human 134591 Alexa Fluor 700 NK phenotyping NK Med
Ki-67 Human B56 Brilliant Blue 660-P2 Proliferation T cell, TCRgd, NK, ILC, MAIT, monocyte Low
CD57 Human HNK-1 Brilliant Violet 711 NK phenotyping NK High
CD127 Human hIL-7R-M21 Brilliant Blue 790-P ILC phenotyping ILC Med
Fc epsilon R1 gamma Human polyclonal FITC NK phenotyping NK Med
CD16 Human 3G8 Brilliant Ultraviolet 395 NK, monocyte, ILC NK, monocyte Med
Dead cells Human Live/Dead Fix Blue Viability
CD3 Human UCHT1 Brilliant Ultraviolet 496 T cell T cell, TCRgd, MAIT High
TCR gamma/delta Human 11F2 Brilliant Violet 421 γδ T cell TCRgd Low
CD314 Human 1D11 Brilliant Violet 480 NK phenotyping TCRgd, NK, T cell Low
CD56 Human HCD56 Brilliant Violet 570 NK phenotyping NK Low
TCR V delta 2 Human B6 Brilliant Violet 605 γδ T cell subset vd2 TCRgd Med
CD38 Human HB-7 Brilliant Violet 650 Activation T cell, monocyte, TCRgd Low
TCR V alpha 7.2 Human 3C10 Brilliant Violet 786 MAIT MAIT Med
CD14 Human MfP9 PerCP-Cy5.5 Monocyte monocyte High
NKp46 Human 9E2/NKp46 PE NK phenotyping NK Low
CD294 Human BM16 PE-CF594 ILC phenotyping ILC Med
CD117 Human 104D2 PE-Cy5.5 ILC phenotyping ILC Med
CD337 Human p30-15 Alexa Fluor 647 NK phenotyping NK Low
CD161 Human HP-3G10 APC-Fire 750 MAIT/ILC phenotyping MAIT, ILC, NK Low
HLA DR Human G46-6 Brilliant Ultraviolet 661 Activation T cell, monocyte, NK Low
CD8a Human SK1 Brilliant Ultraviolet 805 T cell T cell High
CD159a (NKG2A) Human Z199 PE-Cy7 NK phenotyping NK, TCRgd Med
CD19 Human SJ25C1 Brilliant Ultraviolet 563 B cell Low
CD27 Human L128 Brilliant Ultraviolet 737 NK phenotyping T cell, TCRgd Med
CD4 Human SK3 Brilliant Violet 750 T cell T celll High
CD33 Human WM53 PE-Cy5 Granulocyte/monocyte exclusion monocyte High