OMIP‐096: A 24‐color flow cytometry panel to identify and characterize CD4+ and CD8+ tissue‐resident T cells in human skin, intestinal, and type II mucosal tissue

Author:

O'Neil Thomas R.12,Harman Andrew N.12,Cunningham Anthony L.12,Nasr Najla12,Bertram Kirstie M.12ORCID

Affiliation:

1. Centre for Virus Research, The Westmead Institute for Medical Research Westmead Australia

2. The Westmead Clinical School, Faculty of Medicine and Health The University of Sydney Sydney Australia

Abstract

AbstractThere is a great need to understand human immune cells within tissue, where disease manifests and infection occurs. Tissue‐resident memory T cells (TRMs) were discovered over a decade ago, there is a great need to understand their role in human disease. We developed a 24‐color flow cytometry panel to comprehensively interrogate CD4+ and CD8+ TRMs isolated from human tissues. When interrogating cells within human tissue, enzymatic methods used to liberate cells from within the tissue can cause cleavage of cell surface markers needed to phenotype these cells. Here we carefully select antibody clones and evaluate the effect of enzymatic digestion on the expression of markers relevant to the identification of T cell residency, as well as markers relevant to the activation and immunoregulation status of these cells. We have designed this panel to be applicable across a range of human tissues including skin, intestine, and type II mucosae such as the vagina.

Funder

National Health and Medical Research Council

Neil and Norma Hill Foundation

Publisher

Wiley

Subject

Cell Biology,Histology,Pathology and Forensic Medicine

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