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Basophil Activation Test for Allergy Diagnosis
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Basophil Stimulation and Signaling Pathways.

Edward F Knol1, Bernhard F Gibbs2

  • 1Department of Immunology, University Medical Center Utrecht, Utrecht, The Netherlands. e.f.knol@umcutrecht.nl.

Methods in Molecular Biology (Clifton, N.J.)
|August 9, 2020
PubMed
Summary

This study details methods for analyzing human basophil signaling and mediator release. It highlights flow cytometry

Keywords:
BasophilsFlow cytometryIgE receptorsInhibitorsSignalingWestern blotting

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Area of Science:

  • Immunology
  • Cellular Signaling
  • Allergy Research

Background:

  • Intracellular signaling cascades in basophils, crucial for allergic responses, are not fully understood.
  • Analyzing mediators like histamine, LTC4, IL-4, and IL-13 often requires large numbers of purified basophils.
  • Existing methods like Western blotting and RT-PCR have limitations with primary human basophil analysis.

Purpose of the Study:

  • To provide methods for studying mediator release and signaling characteristics in primary human basophils.
  • To address the limitations of current techniques for analyzing basophil function.
  • To enable the study of intracellular signaling proteins and calcium concentrations using flow cytometry.

Main Methods:

  • Utilizing flow cytometry for analyzing intracellular signaling proteins and cytosolic free calcium concentrations.
  • Employing basophil-specific gating strategies for accurate cell analysis.
  • Focusing on primary human basophils to overcome limitations of cell lines.

Main Results:

  • Flow cytometry allows analysis of signaling proteins and calcium levels in low cell numbers.
  • Primary human basophils can be effectively used for studying mediator release and signaling.
  • This approach overcomes challenges associated with traditional methods and cell lines.

Conclusions:

  • Flow cytometry is a valuable tool for studying human basophil signaling and mediator release.
  • The described methods enable detailed analysis of basophil function using primary cells.
  • This research contributes to a better understanding of allergic and inflammatory processes.