A protocol for combining proliferation, tetramer staining and intracellular cytokine detection for the flow-cytometric analysis of antigen specific T-cells

Lydia Tesfa, H. D. Volk, F. Kern

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Flow-cytometry can be used in different ways in order to analyze or enumerate antigen specific T-cells. The three basic principles are direct staining of the T-cell receptor using so called tetramer reagents, staining intracellular cytokines following antigen-specific ex vivo T-cell activation or staining with dyes that are incorporated (increase in staining) or distributed between daughter cells (decrease in staining) upon proliferation in response to a specific antigen challenge. Each system has its advantages and disadvantages. Here we demonstrate that tetramer staining, cytokine flow cytometry and staining with CFDA-SE can be combined permitting the analysis of proliferation and cytokine production with a subset of T-cells specific for a single peptide antigen.

Original languageEnglish (US)
Pages (from-to)366-370
Number of pages5
JournalJournal of Biological Regulators and Homeostatic Agents
Volume17
Issue number4
StatePublished - Oct 2003
Externally publishedYes

Fingerprint

cytokines
T-lymphocytes
Staining and Labeling
Cytokines
antigens
T-Lymphocytes
Antigens
flow cytometry
Flow Cytometry
T-Lymphocyte Subsets
T-Cell Antigen Receptor
staining
dyes
Coloring Agents
peptides
Peptides
receptors
cells

Keywords

  • Antigen-specific T-cells
  • CFDA-SE
  • Cytokine flow cytometry
  • Tetramers

ASJC Scopus subject areas

  • Immunology
  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Physiology (medical)
  • Medicine (miscellaneous)
  • Physiology
  • Agricultural and Biological Sciences(all)

Cite this

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AU - Volk, H. D.

AU - Kern, F.

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AB - Flow-cytometry can be used in different ways in order to analyze or enumerate antigen specific T-cells. The three basic principles are direct staining of the T-cell receptor using so called tetramer reagents, staining intracellular cytokines following antigen-specific ex vivo T-cell activation or staining with dyes that are incorporated (increase in staining) or distributed between daughter cells (decrease in staining) upon proliferation in response to a specific antigen challenge. Each system has its advantages and disadvantages. Here we demonstrate that tetramer staining, cytokine flow cytometry and staining with CFDA-SE can be combined permitting the analysis of proliferation and cytokine production with a subset of T-cells specific for a single peptide antigen.

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