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Multi‐parametric phospho‐flow cytometry: A crucial tool for T lymphocyte signaling studies
Author(s) -
Goldeck David,
Low Ivy,
Shadan Nurhidaya Binte,
Mustafah Seri,
Pawelec Graham,
Larbi Anis
Publication year - 2013
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22252
Subject(s) - signal transduction , biology , flow cytometry , effector , phosphorylation , microbiology and biotechnology , mass cytometry , lymphocyte , population , cell signaling , cell sorting , cd8 , computational biology , immunology , immune system , genetics , medicine , gene , environmental health , phenotype
Tools such as protein immunoblotting have proven benefits for investigating T lymphocyte signaling but have several drawbacks such as the number of cells required and the difficulty of distinguishing subset‐specific differences without expensive and invasive cell sorting. Recent advances in immunology and the identification of T lymphocyte sub‐populations making up only a very small fraction of the total population highlight the importance of studying signaling in those small subsets in a feasible, cost‐effective, high‐throughput manner. To this end, we have developed a simplified protocol to study both intracellular phosphorylation patterns of important signal transduction molecules concomitantly with T cell surface marker expression. A multi‐parametric analysis may allow the quantification of the phosphorylation of up to five signaling molecules in CD4 and CD8 T lymphocytes and their naïve, central memory, effector memory, and TEMRA subsets. This enables precise identification of subset‐specific signaling and alterations of signaling pathways in physiological and pathological situations. The importance of such detailed analysis is discussed. © 2013 International Society for Advancement of Cytometry