OMIPs Directory

OMIP-056

Evaluation of Human Conventional T Cells, Donor‐Unrestricted T Cells, and NK Cells Including Memory Phenotype by Intracellular Cytokine Staining

Cytometry PART A, Volume 95, Issue 7, 722-725 (2019)
One Dintwe, Shamiska Rohith, Katharine V. Schwedhelm, M. Juliana McElrath, Erica Andersen‐Nissen, Stephen C. De Rosa

Description: A 26‐color staining panel was developed to profile human antigen‐specific T cells in an intracellular cytokine staining (ICS) assay using peptide pools to various antigens of interest. In addition to multiple functional markers, the panel includes differentiation/activation markers and markers to assess γδ, mucosal‐associated invariant T, and NK T cells as well as conventional NK cells. Panel optimization was performed using previously cryopreserved PBMC from healthy adults, and then, expression of key functional markers in the panel was cross‐validated against a validated ICS assay used in the HIV Vaccine Trials Network (HVTN). The panel is currently being used to evaluate the responses to tuberculosis and malaria vaccine candidates in volunteers from different geographic areas.

Species: Human

Tissue: Peripheral blood

Cell Subset: Ctl, Mait, T helper, T memory, Tcrgd, Nk, Nkt

Keyword: Antigen-specific t cell response, Cryopreserved pbmc, Hiv, Malaria, Tuberculosis (tb)

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
Dead cells Human Live/Dead Fix Blue Viability
CD45RA Human HI100 Brilliant Ultraviolet 496 Differentiation T memory, T helper, CTL, TCRgd Med
KLRG1 Human REA261 PE-Vio615 Differentiation T memory, NK Low
CD14 Human MfP9 Brilliant Blue 660-P2 Monocyte Monocyte High
IFN-gamma Human B27 V450 Function T helper, CTL, NK, MAIT, TCRgd High
CD3 Human UCHT1 Brilliant Ultraviolet 395 T cell lineage TCRgd, CTL, T helper, MAIT High
CD56 Human CMSSB PE-Cy5.5 NK,NKT NK Low
IL-4 Human MP4-25D2 APC Function T helper Low
Perforin Human B-D48 FITC Function CTL, MAIT, NK Med
IL-22 Human 22URTI PE Function T helper Low
CD183 Human 1C6/CXCR3 PE-Cy5 T helper T helper, CTL Med
CCR7 Human G043H7 Brilliant Violet 785 Differentiation T memory, T cell, TCRgd Low
CD40L Human 24-31 PE-Cy7 Function T memory, CTL, T helper, TCRgd Med
CD8a Human RPA-T8 Brilliant Ultraviolet 563 T cell lineage CTL High
TNF Human Mab11 Brilliant Violet 750 Function CTL, T helper Med
TCR V alpha 7.2 Human REA179 APC-Vio770 MAIT MAIT Med
CD26 Human M-A261 Brilliant Violet 711 MAIT MAIT Med
CD16 Human 3G8 Brilliant Violet 570 NK,NKT NK Med
CD161 Human DX12 Brilliant Violet 650 MAIT MAIT, NK Low
HLA DR Human G46-6 Brilliant Ultraviolet 661 Activation T helper, T memory, CTL, NK, Monocyte Low
Granzyme A Human CB9 Alexa Fluor 700 Function CTL, MAIT, NK Med
TCR gamma/delta Human 11F2 Brilliant Violet 480 TCRgd TCRgd Low
IL-17A Human N49-653 Brilliant Ultraviolet 737 Function T helper, TCRgd Low
IL-2 Human MQ1-17H12 Brilliant Blue 700 Function CTL, T helper, NK, TCRgd Med
CD4 Human RPA-T4 Brilliant Ultraviolet 805 T cell lineage T helper High
IL-13 Human JES10-5A2 APC Function T helper Low
CCR6 Human 11A9 Brilliant Violet 605 T helper T helper, CTL Med