OMIPs Directory

OMIP-058

30‐Parameter Flow Cytometry Panel to Characterize iNKT, NK, Unconventional and Conventional T Cells

Cytometry PART A, Volume 95, Issue 9, 946-951 (2019)
Thomas Liechti, Mario Roederer

Description: This panel enables the in‐depth immunophenotyping of unconventional CD3+ T‐cell populations such as invariant natural killer T cells (iNKT; CD1d:PBS57 tetramer), γδ T cell subsets (TCR Vδ1, TCR Vδ2, TCR Vγ9), and mucosal‐associated invariant T cells (MAIT; CD161, TCR Vα7.2) in combination with conventional CD3+ T cells (CD4, CD8) and natural killer (NK) cell subsets (CD16, CD56). We included markers to assess differentiation stages (CCR7, CD27, CD28, CD45RA, CD57, and CD95) and expression of cytokine receptors (CD122, CD127), activation markers (CD38, HLA‐DR), chemokine receptors (CCR5, CXCR5), the co‐inhibitory molecule PD‐1, and activating and inhibitory NK receptors (CD158, CD244, and CD318). This unique combination of markers enables a thorough phenotypic characterization of these cell subsets.

Species: Human

Tissue: Peripheral blood

Cell Subset: T cell, Mait, T memory, Tcrgd, Treg, Nk, Nkt

Keyword: Activation, Immune checkpoint

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CD57 Human NK-1 Brilliant Blue 630-P2 Immune senescence activated T, NKT, NK, γδ T cell High
CD244 Human 25235 Brilliant Blue 660-P2 Co-stimulatory/inhibitory receptor NK, activated T Low
TCR V delta 2 Human B6 PE-CF594 γδ T cell γδ T cell Med
PD-1 Human EH12.1 Brilliant Ultraviolet 661 Inhitibory receptor activated T, γδ T cell Low
CD38 Human HB7 Brilliant Violet 750 n/a activated T Low
CCR7 Human 150503 Brilliant Ultraviolet 395 Differentiation T memory, γδ T cell Low
CD1d Human tetramer APC iNKT iNKT Unspecified
Dead cells Human Live/Dead Fix Blue viability
TCR V delta 1 Human TS8.2 FITC γδ T cell γδ T cell Low
CD127 Human hIL-7R-M21 Brilliant Blue 700 Cytokine receptor T memory, γδ T cell, MAIT Med
TCR V gamma 9 Human B3 PE γδ T cell γδ T cell Med
CD161 Human DX12 PE-Cy5 MAIT MAIT Low
CD314 Human 1D11 PE-Cy7 KIR/NKG2D NK, activated T, NKT Low
CD45RA Human HI100 Alexa Fluor 700 Differentiation T memory, γδ T cell Med
CD4 Human SK3 Brilliant Ultraviolet 805 CD4 T activated T High
CD122 Human Mik-b3 Brilliant Violet 421 Cytokine receptor NK, γδ T cell Low
CD158 Human DX27 Brilliant Violet 605 KIR/NKG2D NK Med
CD28 Human CD28.2 Brilliant Violet 650 Differentiation T memory, γδ T cell Med
TCR V alpha 7.2 Human 3C10 Brilliant Violet 711 MAIT MAIT Med
CD56 Human NCAM16.2 Brilliant Ultraviolet 563 NK NK, NKT Low
CD8a Human RPA-T8 Brilliant Violet 570 CD8 T activated T High
CD185 Human RF8B2 Brilliant Blue 790-P Chemokine receptor γδ T cell, activated T Low
HLA DR Human Tu36 PE-Cy5.5 Activation activated T Low
CD95 Human Dx2 Brilliant Ultraviolet 737 Differentiation activated T Low
CD3 Human UCHT1 Brilliant Violet 510 CD3 T γδ T cell, activated T, MAIT, iNKT High
CD16 Human 3G8 Brilliant Ultraviolet 496 NK NK Med
CD27 Human L128 Brilliant Violet 786 Differentiation T memory, γδ T cell Med
CCR5 Human 2D7/CCR5 APC-Cy7 Chemokine receptor activated T, NK Med