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Author Spotlight: Investigating the Pathophysiology of Eosinophilic Esophagitis

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Assessing Phenotypic Heterogeneity in Intestinal Tissue Eosinophils.

Courtney L Olbrich^1,2, Leigha D Larsen^1,2, Lisa A Spencer^3,4

¹Section of Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, University of Colorado School of Medicine, Aurora, CO, USA.

Methods in Molecular Biology (Clifton, N.J.)

|January 24, 2021

Summary

Flow cytometry now identifies tissue eosinophils in more organs. This study details a new protocol for isolating mouse intestinal eosinophils, applicable to human tissues too.

Keywords:

Eosinophil heterogeneity Eosinophils Intestinal leukocytes Leukocyte isolation Surface phenotype

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Area of Science:

Immunology
Cell Biology

Background:

Eosinophils are primarily tissue-resident leukocytes.
Flow cytometry is increasingly used to study tissue-resident immune cells.
Understanding tissue eosinophil heterogeneity is crucial in health and disease.

Purpose of the Study:

To describe a novel tissue digestion protocol for isolating eosinophils from the mouse small intestine.
To establish a flow cytometry gating strategy for identifying and characterizing tissue eosinophils.
To demonstrate the protocol's adaptability for colonic and human intestinal eosinophils.

Main Methods:

Tissue digestion of the mouse small intestine.
Flow cytometry analysis for eosinophil identification.
Development of a specific gating strategy for tissue eosinophils.

Main Results:

Successful isolation and identification of eosinophils from digested mouse small intestine.
Established a reliable flow cytometry gating strategy for tissue eosinophils.
Demonstrated protocol applicability to mouse colonic tissue and human intestinal samples.

Conclusions:

The described protocol enables robust identification and isolation of tissue eosinophils.
This method facilitates deeper phenotypical and functional characterization of tissue-resident eosinophils.
The protocol is adaptable for studying intestinal eosinophils across species and conditions.