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Flow Cytometry Purification of Mouse Meiotic Cells
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Comprehensive Phenotyping of T Cells Using Flow Cytometry.

Charlotte M Mousset1, Willemijn Hobo1, Rob Woestenenk1

  • 1Department of Laboratory Medicine - Laboratory of Hematology, Radboud Institute of Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|February 5, 2019
PubMed
Summary

This review details how to identify diverse T cell subsets, including CD4+ and CD8+ T-helper (Th) and cytotoxic T lymphocyte (Tc) cells, and their memory states using flow cytometry. It provides a guide for comprehensive T cell phenotyping.

Keywords:
T cellT cell subsetdifferentiationflow cytometryphenotyping

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Area of Science:

  • Immunology
  • Cell Biology
  • Flow Cytometry

Background:

  • T cells are crucial for immune defense against pathogens and cancer, but also regulate immune responses to prevent autoimmunity.
  • T cell functions vary widely based on their differentiation and maturation states.
  • Accurate identification of T cell subsets is essential for understanding immune responses.

Purpose of the Study:

  • To provide an overview of methods for identifying various CD4+ and CD8+ T-αβ cell subsets and their effector memory statuses.
  • To discuss the use of flow cytometry for discriminating these T cell subsets based on extracellular markers and intracellular staining.
  • To cover the identification of rare T cell populations and technical considerations in flow cytometry.

Main Methods:

  • Utilizing flow cytometry for the identification of CD4+ T-helper (Th) cells (Th1, Th2, Th9, Th17, Th22, regulatory T cells) and CD8+ cytotoxic T lymphocyte (Tc) cells (Tc1, Tc2, Tc9, Tc17, regulatory T cells).
  • Discriminating T cell subsets through selective extracellular marker staining combined with intracellular transcription factor and/or cytokine analysis.
  • Employing flow cytometry techniques for the identification of antigen-specific T cells and other rare subsets.

Main Results:

  • Established protocols for distinguishing major T cell subsets (Th1-22, Tc1-17, regulatory T cells) and their memory phenotypes (naïve, stem cell memory, central memory, effector memory, effector).
  • Demonstrated the utility of combining extracellular and intracellular staining for precise T cell subset identification.
  • Highlighted technical considerations for flow cytometry, enabling the analysis of even very small T cell populations.

Conclusions:

  • The presented overview offers a comprehensive approach to phenotyping T cell subsets using flow cytometry.
  • This guide facilitates the detailed characterization of diverse T cell populations, crucial for immunological research.
  • Accurate T cell subset identification is key to understanding immune regulation, defense mechanisms, and potential therapeutic targets.