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Pitfalls in the characterization of circulating and tissue-resident human γδ T cells

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Nicola Beucke, Daniela Wesch, Hans-Heinrich Oberg, Christian Peters, Jonas Bochem, Benjamin Weide, Claus Garbe, Graham Pawelec, Susanne Sebens, Christoph Röcken, Hisayoshi Hashimoto, Markus W Löffler, Paola Nocerino, Shahram Kordasti, Dieter Kabelitz, Karin Schilbach, Kilian Wistuba-Hamprecht

Original languageEnglish
Pages (from-to)1097-1105
Number of pages9
JournalJournal of Leukocyte Biology
Volume107
Issue number6
Early online date22 Jan 2020
DOIs
Accepted/In press6 Jan 2020
E-pub ahead of print22 Jan 2020
Published1 Jun 2020

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©2020 Society for Leukocyte Biology.

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Abstract

Dissection of the role and function of human γδ T cells and their heterogeneous subsets in cancer, inflammation, and auto-immune diseases is a growing and dynamic research field of increasing interest to the scientific community. Therefore, harmonization and standardization of techniques for the characterization of peripheral and tissue-resident γδ T cells is crucial to facilitate comparability between published and emerging research. The application of commercially available reagents to classify γδ T cells, in particular the combination of multiple Abs, is not always trouble-free, posing major demands on researchers entering this field. Occasionally, even entire γδ T cell subsets may remain undetected when certain Abs are combined in flow cytometric analysis with multicolor Ab panels, or might be lost during cell isolation procedures. Here, based on the recent literature and our own experience, we provide an overview of methods commonly employed for the phenotypic and functional characterization of human γδ T cells including advanced polychromatic flow cytometry, mass cytometry, immunohistochemistry, and magnetic cell isolation. We highlight potential pitfalls and discuss how to circumvent these obstacles.

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