OMIP-084: 28-color full spectrum flow cytometry panel for the comprehensive analysis of human γδ T cells

Cytometry A. 2022 Oct;101(10):856-861. doi: 10.1002/cyto.a.24564. Epub 2022 May 6.

Abstract

Using full spectrum flow cytometry, we designed a 28-color panel for the analysis of markers known to be associated with the γδ T cell immune response. This panel allows the classification of γδ T cell subsets via specific V gene usage (Vγ9, Vδ1, Vδ2, and Vδ3) of their T cell receptor (TCR) and according to their functional differentiation. Phenotypical surface receptors to distinguish different stages of cell maturation included CD45RA, CD27, CD28, CD127, CD57, and CD16; chemokine receptors CXCR6, CCR5, CCR6, and CX3CR1; NK-associated markers NKG2A, NKG2D, CD56, and CD161, checkpoint-inhibitor PD-1, and activating receptors CD38 and CD25. T cell lineage markers for the analysis of αβ T cells (CD4 and CD8) and MAIT cells (Vα7.2) were also included. This optimized multicolor panel allows a comprehensive immune-profiling of all main human γδ T cell subsets and is suitable for longitudinal or exploratory analysis of γδ T cell development and γδ T cell dynamics in clinical cohorts.

Keywords: Immunophenotyping; MAIT; T cells; cord blood; full spectrum flow cytometry; human PBMCs; γδ T cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD28 Antigens* / metabolism
  • Flow Cytometry
  • Humans
  • NK Cell Lectin-Like Receptor Subfamily K / metabolism
  • Programmed Cell Death 1 Receptor / metabolism
  • Receptors, Antigen, T-Cell, gamma-delta* / metabolism
  • Receptors, Chemokine / metabolism
  • T-Lymphocyte Subsets

Substances

  • CD28 Antigens
  • NK Cell Lectin-Like Receptor Subfamily K
  • Programmed Cell Death 1 Receptor
  • Receptors, Antigen, T-Cell, gamma-delta
  • Receptors, Chemokine