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Related Concept Videos

T Cell Types and Functions01:24

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Updated: Apr 15, 2026

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry
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Quantifying helper cell function of human TFH cells in vitro.

Nina Chevalier1

  • 1Department of Rheumatology and Clinical Immunology, University Freiburg Medical School, Hugstetter Str. 55, Freiburg, 79106, Germany, nina.chevalier@uniklinik-freiburg.de.

Methods in Molecular Biology (Clifton, N.J.)
|April 4, 2015
PubMed
Summary

This study details methods for isolating T follicular helper (TFH)-like cells from blood and tonsils. Researchers can now quantify their capacity to support B cell differentiation and immunoglobulin production.

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Area of Science:

  • Immunology
  • Cellular Biology
  • Hematology

Background:

  • T follicular helper (TFH) cells are crucial for adaptive immunity, supporting B cell responses within germinal centers.
  • Circulating CXCR5(+) CD4(+) T cells are considered precursors or subsets of TFH cells, playing a role in B cell help.
  • Understanding TFH cell function is vital for studying humoral immunity and developing immunotherapies.

Purpose of the Study:

  • To provide detailed protocols for isolating and characterizing TFH-like cells from peripheral blood and tonsils.
  • To establish methods for quantifying the B cell helper function of these TFH-like cells.
  • To enable indirect assessment of TFH cell activity through cytokine and molecule expression analysis.

Main Methods:

  • Isolation of TFH-like cells (CXCR5(+) CD4(+) T cells) from peripheral blood and tonsillar tissues.
  • Co-culture systems involving TFH-like cells and B cells to assess B cell differentiation and immunoglobulin secretion.
  • Quantification of plasma cell differentiation and immunoglobulin production as readouts of B cell help.
  • Indirect assessment via measurement of B cell helper cytokines and TFH-associated molecules (e.g., co-stimulatory molecules).

Main Results:

  • Established protocols allow for the isolation of functional TFH-like cells from human samples.
  • Co-culture experiments successfully demonstrated the capacity of TFH-like cells to induce B cell immunoglobulin secretion and plasma cell differentiation.
  • Indirect measurement of specific cytokines and molecules provides a viable alternative for assessing TFH cell helper function.

Conclusions:

  • The described methods provide a robust framework for studying TFH cell biology and function in vitro.
  • These techniques are essential for investigating the role of TFH cells in various immunological settings, including autoimmune diseases and vaccine responses.
  • Quantifying B cell helper function is critical for understanding humoral immunity and developing targeted B cell therapies.