OMIPs Directory

OMIP-036

Co-inhibitory receptor (immune checkpoint) expression analysis in human T cell subsets

Cytometry PART A, Volume 89, Issue 10, 889–892 (2016)
Zachary R. Healy, David M. Murdoch

Description: This panel was optimized to quantify inhibitory receptor expression on CD4 and CD8 T cells from differentiation and activation subsets. Six inhibitory (i.e., immune checkpoint) receptors (PD-1, TIM-3, LAG-3, CD160, BTLA, CTLA-4) were chosen based upon previously published observations suggesting their role in modulating CD4 and CD8 T cell activation in response to persistent antigen exposure. Furthermore, given the important observations that inhibitory receptor expression varies by differentiation and prior antigen experience, markers of T cell differentiation status and prior antigen experience (CCR7, CD45RA, CD28, CD127, KLRG1) were also included. This panel was developed and optimized for use in cryopreserved human peripheral blood mononuclear cells (PBMCs), although it has also been applied in fresh PBMCs as well as other bodily fluids (e.g., malignant ascites).

Species: Human

Tissue: Ascites, Peripheral blood

Cell Subset: T cell, T helper, T memory, Ctl, Treg

Keyword: Cryopreserved pbmc, Immune checkpoint

Machine: BD Fortessa

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
LAG-3 Human 3DS223H PE-Cy7 Inhibitory Receptors T cell Low
CD8a Human SK1 Brilliant Ultraviolet 805 Phenotyping T cell, T memory High
Dead cells Human Zombie Aqua Viability
CD14 Human M5E2 Brilliant Violet 510 Dump/exclusion High
CD3 Human SK7 Brilliant Ultraviolet 395 Phenotyping T cell, T memory High
CD4 Human SK3 Brilliant Ultraviolet 496 Phenotyping T cell, T memory High
CD45RA Human HI100 Brilliant Blue 515 Maturation/Activation T cell, T memory Med
CD28 Human CD28.2 APC-H7 Maturation/Activation T cell, T memory Med
KLRG1 Human SA231A2 Alexa Fluor 647 Maturation/Activation T cell, T memory Low
PD-1 Human EH12.2H7 Brilliant Violet 711 Inhibitory Receptors T cell Low
TIM-3 Human F38-2E2 Brilliant Violet 605 Inhibitory Receptors T cell Low
CD160 Human BY55 PerCP-Cy5.5 Inhibitory Receptors T cell Low
CD272 Human MIH26 Brilliant Violet 421 Inhibitory Receptors T cell Low
CTLA-4 Human BNI3 PE-CF594 Inhibitory Receptors T cell Low
CCR7 Human G043H7 Brilliant Violet 785 Maturation/Activation T cell, T memory Low
CD127 Human A019D5 Brilliant Violet 650 Maturation/Activation T cell, T memory Med
CD19 Human HIB19 Brilliant Violet 510 Dump/exclusion Low