OMIPs Directory

OMIP-071

A 31-Parameter Flow Cytometry Panel for In-Depth Immunophenotyping of Human T-Cell Subsets Using Surface Markers

Cytometry PART A, Volume 99, Issue 3, 273-277 (2021)
Song-Rong Wang, Na Zhong, Xin-Mei Zhang, Zhi-Bin Zhao, Robert Balderas, Liang Li, Zhe-Xiong Lian

Description: Dissecting the functional diversity of T cells is critical in elucidating mechanisms and in developing therapies for various diseases. Here, we designed a 31-parameter (29-color) panel to enable the characterization of T cell subsets and immunophenotyping of the human peripheral blood and lymph nodes using cell surface staining. In addition to adaptive T-cell markers, TCR Vα24-Jα18, TCR γδ, TCR Vɑ7.2, and CD161 were included to identify iNKT, γδ T, and MAIT cells, respectively, which are innate-like T cells. C-X-C chemokine receptors (CXCR3, CXCR4, CXCR5, CXCR6) and C-C motif chemokine receptors (CCR4, CCR6, CCR7) were included to enable the identification of Th cell subsets (Th1, Th2, Th17), Tfh cell subsets (Tfh1, Tfh2, Tfh17), and Th cells with specific homing capacities. Furthermore, in this panel, we also used markers for assessing cell differentiation (CD45RO, CD7), activation (CD57, CD95, HLA-DR) and the expression of some co-signaling molecules (PD-1, NKG2D, CD28). Particularly, CD69 and CD103 were included for the further analysis of tissue resident memory T (Trm) cells. This panel would enable the in-depth immunophenotyping of human T-cell subsets, and may be applied in the monitoring, prognosis, and mechanistic studies of various immune-related diseases.

Species: Human

Tissue: Lymph node, Peripheral blood

Cell Subset: T cell, Ctl, Mait, T helper, T memory, Tcrgd, Th1, Th17, Th2, Tissue resident memory (trm), Treg, Nk, Nkt, Tfh

Keyword: Activation, Immune checkpoint

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CD185 Human RF8B2 Brilliant Ultraviolet 615 Tfh marker Tfh Low
TCR V alpha 7.2 Human OF-5A12 Brilliant Violet 570 MAIT cell lineage MAIT Med
CD103 Human 2E7 Brilliant Blue 630-P2 Tissue residency marker CTL, T helper Med
CD314 Human 1D11 Brilliant Blue 660-P2 Costimulation molecule NK, NKT, TCRgd, CTL Low
CCR7 Human 2-L1-A APC-Cy7 Naive cell marker Thelper, CTL, TCRgd Low
CD194 Human 1G1 Brilliant Ultraviolet 395 Th marker Thelper Med
CD127 Human hIL-7R-M21 PE-Cy5 Regulatory T cell lineage Treg Low
CD28 Human CD28.2 PE Costimulation molecule CTL, MAIT, NKT, TCRgd, T helper Med
Dead Cell All Species Zombie UV viability
CD7 Human M-T701 PE-Cy7 Naive/effector T cell T helper, CTL, TCRgd, NKT, MAIT Med
CD25 Human 2A3 Brilliant Violet 421 Regulatory T cell lineage Treg Low
CD45 Human HI30 APC-R700 White blood cell lineage TCRgd, T helper, CTL, NKT, NK, MAIT High
CD45RO Human UCHL1 PE-CF594 Memory cell marker T helper, CTL, NKT, MAIT, TCRgd Med
CD161 Human DX12 Brilliant Ultraviolet 737 MAIT cell marker MAIT, NK, NKT Low
PD-1 Human EH12.1 Brilliant Blue 515 Exhaustion marker CTL, NKT, MAIT Low
CD183 Human G025H7 Brilliant Violet 510 Tfh/Th marker CTL, T helper Low
CXCR4 Human 12G5 Brilliant Ultraviolet 563 Th marker CTL, T helper Low
CD186 Human 13B 1E5 Brilliant Blue 700 Th marker CTL, T helper Low
TCR V alpha 24 J alpha 18 Human 6B11 Brilliant Violet 711 iNKT cell marker NKT Med
HLA DR Human G46-6 Brilliant Violet 605 Activation marker TCRgd Med
CD8a Human RPA-T8 Brilliant Violet 786 T cell lineage CTL, NKT, MAIT, TCRgd High
CCR6 Human 11A9 Brilliant Ultraviolet 496 Tfh/Th marker CTL, T helper Med
CD95 Human Dx2 Brilliant Ultraviolet 395 Activation/exhaustion marker CTL, T helper Low
CD57 Human NK-1 Brilliant Violet 480 Terminal Differentiation NKT, NK, TCRgd, CTL High
TCR delta/gamma Human B1 APC γδ T cell lineage TCRgd Low
CD56 Human NCAM16.2 Brilliant Violet 650 NK cell lineage/NKT-like cell NK, NKT Low
CD3 Human UCHT-1 Brilliant Ultraviolet 805 T cell lineage NKT, CTL, T helper, Treg, TCRgd High
CD4 Human SK3 Brilliant Violet 750 T cell lineage NKT, Treg, T helper, TCRgd High
CD69 Human FN50 Brilliant Blue 790-P Activation/ tissue residency marker CTL, T helper, NK, NKT Med