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Flow Cytometric Measurement Of ROS Production In Macrophages In Response To FcγR Cross-linking

Flow Cytometric Measurement Of ROS Production In Macrophages In Response To FcγR Cross-linking

Published on: March 7, 2019

Eosinophils oxidize damage-associated molecular pattern molecules derived from stressed cells.

Ramin Lotfi¹, Gloria Isabelle Herzog, Richard Anthony DeMarco

¹Institut für klinische Transfusionsmedizin und Immungenetik Ulm, Institut für Transfusionsmedizin Universitat Ulm, Ulm, Germany.

Journal of Immunology (Baltimore, Md. : 1950)

|October 2, 2009

Summary

Eosinophils detect necrotic tumor material, releasing potent molecules. This study suggests eosinophils may help clear dead cells through oxidation, potentially informing new cancer therapies.

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Area of Science:

Immunology
Cell Biology
Oncology

Background:

Eosinophils (Eos) accumulate in tumor necrotic areas.
Necrotic cell debris contains damage-associated molecular pattern (DAMP) molecules.

Purpose of the Study:

To investigate eosinophil responses to necrotic material and DAMPs.
To explore the role of High Mobility Group Box 1 (HMGB1) in eosinophil activation.
To assess the therapeutic potential of eosinophil-mediated oxidative degradation of necrotic debris.

Main Methods:

Stimulation of eosinophils with cell lysates and purified HMGB1.
Measurement of eosinophil degranulation (MBP, EPO release) and oxidative burst.
Assessment of eosinophil survival and chemotaxis.
Inhibition studies using anti-receptor for advanced glycation end product (RAGE) antibodies.

Main Results:

Necrotic cell lysates and HMGB1 induced eosinophil degranulation and enhanced oxidative burst.
Oxidation of cell lysates diminished their stimulatory capacity.
HMGB1 increased eosinophil survival and acted as a chemoattractant.
Eosinophils express RAGE, and anti-RAGE antibodies blocked HMGB1-mediated effects.
Eosinophils showed heightened sensitivity to HMGB1 and necrotic material, particularly in peroxide generation.

Conclusions:

Eosinophils can sense necrotic cell death and migrate to injury sites.
Eosinophil-mediated oxidative degradation of necrotic material may modulate immunity in tumors.
Targeting HMGB1 or promoting oxidative denaturation presents novel therapeutic strategies for cancer.