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Analysis of Th1/Th2 T-cell subsets

Alla Skapenko1, Hendrik Schulze-Koops

  • 1Nikolaus Fiebiger Center for Molecular Medicine, Clinical Research Group III, University of Erlangen-Nuremberg, Erlangen, Germany.

Methods in Molecular Medicine
|November 7, 2007
PubMed

Insights

Understanding T-helper (Th) cell subsets, crucial for immune responses, involves analyzing their signature cytokines like interferon-gamma and interleukin-4. This study details flow cytometry for detecting these cytokines in human Th cells.

Area of Science:

  • Immunology
  • Cellular Biology

Background:

  • Specific immune responses rely on activated CD4+ T-helper (Th) cells.
  • Two main subsets, Th1 and Th2, are defined by their distinct cytokine profiles: interferon-gamma (Th1) and interleukin-4 (Th2).
  • Analyzing these signature cytokines is key to understanding immune responses, both protective and pathogenic.

Purpose of the Study:

  • To review methods for analyzing Th cell subsets based on cytokine production.
  • To present a detailed protocol for intracellular cytokine detection in human Th subsets using flow cytometry.

Main Methods:

  • The study discusses various techniques including reverse transcriptase polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), ELISpot, and intracellular flow cytometry.
  • A standard protocol for flow cytometric analysis of cytoplasmic cytokines in human Th subsets is described.

Main Results:

  • Flow cytometry offers a robust method for identifying and quantifying Th1 and Th2 cells based on their cytokine production.
  • The described protocol enables detailed analysis of human Th subsets.

Conclusions:

  • Accurate analysis of Th cell subsets is vital for advancing the understanding of immune mechanisms.
  • Flow cytometry provides a valuable tool for dissecting Th cell-mediated immunity.

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