OMIPs Directory

OMIP-070

NKp46‐Based 27‐Color Phenotyping to Define Natural Killer Cells Isolated From Human Tumor Tissues

Cytometry PART A, Volume 97, Issue 10, 1052-1056 (2020)
Marie Frutoso, Florian Mair, Martin Prlic

Description: This 27‐color panel has been validated and optimized to comprehensively profile natural killer (NK) cells isolated from human tumors using a collagenase Type II‐based digestion protocol. We confirmed that detection of protein expression by antibodies used in our final panel was not affected during tissue digestion. During this evaluation process, we found that detection of CD56, a biomarker typically used to identify NK cells, was affected substantially by collagenase‐based digestion. Thus, our panel is centered around expression of NKp46, which is sufficient to identify NK cells and not affected by the tissue collagenase digestion step. Our panel further includes biomarkers used to extrapolate NK‐cell maturation, differentiation, migration, homing potential, and functional state. Our panel is intended to provide in‐depth characterization of human NK cells isolated from tissues, which we specifically tested using oral squamous cell carcinomas tissues, but it is compatible with other tissues that can be dissociated with a collagenase Type II‐based protocol.

Species: Human

Tissue: Peripheral blood, Tumor

Cell Subset: Nk

Keyword: Cell migration, Collagenase digestion, Oral squamous cell carcinoma

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
Dead cells Human Live/Dead Fix Blue viability
Streptavidin Human Brilliant Blue 630-P2 secondary reagent to NKp46 NK Unspecified
CD38 Human HIT2 Brilliant Blue 790-P activation NK, T cell Low
CD14 Human M5E2 Brilliant Violet 570 monocyte High
CD3 Human UCHT1 Brilliant Ultraviolet 661 T cell T cell High
CD16 Human 3G8 Brilliant Ultraviolet 496 NK and ADCC NK Med
CD314 Human 1D11 PE-Cy7 activation NK Low
CD244 Human 25235 Brilliant Violet 421 activation NK, T cell Low
CD161 Human DX12 Brilliant Blue 700 inhibitory NK, T cell Med
TIGIT Human 741182 Brilliant Violet 711 co-inhibitory NK, T cell Low
CD69 Human FN50 Brilliant Violet 650 residency and activation NK, T cell Med
CD39 Human TU66 PE-CF594 residency and activation T cell, NK Low
CD11b Human ICRF44 PE-Cy5 maturation and adhesion NK, T cell Low
CX3CR1 Human 2A9-1 Brilliant Ultraviolet 737 migration NK, T cell Low
CD103 Human Ber-ACT8 Brilliant Violet 750 residency T cell, NK Med
Ki-67 Human SolA15 eFluor 660 proliferation NK, T cell Low
CD25 Human 2A3 Brilliant Ultraviolet 563 activation T cell Med
CD56 Human CMSSB PE-Cy5.5 activation, NK for PB NK Low
HLA DR Human L243 APC-H7 exclusion/activtion Med
CD19 Human SJ25C1 (also known as SJ25-C1) Brilliant Ultraviolet 395 B cell Low
NKp46 Human 900 NK and activation NK Low
CD159a (NKG2A) Human REA110 PE inhibitory NK Low
CD27 Human M-T271 Brilliant Blue 660-P2 maturation T cell Low
CD45 Human HI30 Brilliant Ultraviolet 805 lymphocyte T cell, NK High
CD127 Human hIL-7R-M21 Brilliant Violet 786 ILC T cell Med
CD2 Human RPA-2.10 Brilliant Violet 605 co-stimulatory NK, T cell Med
CD57 Human NK-1 FITC maturation and differentiation NK High
Granzyme B Human QA16A02 Alexa Fluor 700 cytolytic ability T cell, NK Med