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OMIP-046

Characterization of invariant T cell subset activation in humans

Kerri G. Lal, Edwin Leeansyah, Johan K. Sandberg, Michael A. Eller Cytometry PART A, Volume 93, Issue 5, 499-503 (2018)

PURPOSE: This panel was developed to characterize the two most commonly studied invariant T cell subsets: mucosal‐associated invariant T (MAIT) cells and invariant natural killer T (iNKT) cells in humans and measure their expression of exhaustion and activation markers. The panel was optimized and designed for application in cryopreserved peripheral blood mononuclear cells (PBMC) (Table 1). Designed to be used to measure frequency and phenotype of invariant T cell subtypes in acute HIV‐1 infection, optimization was performed on chronically HIV‐1 infected and uninfected donors and can be applied to a variety of human disease cohorts.

CELL TYPE: PBMC

MACHINE: BD LSR2 SORP

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua Viability
TCR V alpha 24 Human C15 FITC iNKT
CD8 Human SK1 PerCP-Cy5.5 T cell lineage
CD57 Human hCD57 Pacific Blue Activation
CD161 Human HP-3G10 Brilliant Violet 605 MAIT
HLA-DR Human L243 Brilliant Violet 650 Activation
CD38 Human HIT2 Brilliant Violet 711 Activation
PD-1 Human EH12.1 Brilliant Violet 786 Exhaustion
TCR V alpha 7.2 Human 3C10 APC MAIT
CD14 Human M5E2 Alexa Fluor 700 Exclusion
CD19 Human Alexa Fluor 700 Exclusion
CD4 Human SK3 APC-H7 T cell lineage
TCR V beta 11 Human C21 PE iNKT
CD3 Human S4.1 PE-Texas Red T cell lineage
TIGIT Human MBSA43 PE-Cy7 Exhaustion