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Related Experiment Video

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Basophil Activation Test for Allergy Diagnosis
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Basophil stimulation and signaling pathways.

Edward F Knol1, Bernhard F Gibbs

  • 1Department Immunology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands, e.f.knol@umcutrecht.nl.

Methods in Molecular Biology (Clifton, N.J.)
|August 24, 2014
PubMed
Summary

This study details methods for analyzing primary human basophils, focusing on mediator release and signaling pathways. Flow cytometry offers a sensitive approach for studying these crucial immune cells.

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Area of Science:

  • Immunology
  • Cellular Biology
  • Molecular Biology

Background:

  • Intracellular signaling cascades in primary basophils are not well understood.
  • Analyzing inflammatory mediators like histamine, LTC4, IL-4, and IL-13 requires specific methods.
  • Traditional methods often need large numbers of purified basophils.

Purpose of the Study:

  • To outline methods for studying mediator releases and signaling in primary human basophils.
  • To highlight the utility of flow cytometry for analyzing low cell numbers.
  • To address limitations in extrapolating data from cell lines.

Main Methods:

  • Utilizing primary human basophils.
  • Employing flow cytometry for analyzing intracellular signaling proteins and calcium concentrations.
  • Investigating mediator releases and gene expression.

Main Results:

  • Flow cytometry enables analysis of signaling proteins and calcium in low cell numbers with basophil-specific gating.
  • Established protocols for studying mediator release and signaling characteristics in primary human basophils.

Conclusions:

  • Primary human basophils are essential for understanding basophil signaling and mediator release.
  • Flow cytometry provides a valuable tool for analyzing functional characteristics of limited primary basophil samples.
  • This chapter provides a practical guide for researchers in the field.