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Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions
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The influence of extracellular tissue on neutrophil function and its possible linkage to inflammatory diseases

Richard F Kraus1, Michael A Gruber1, Martin Kieninger1

  • 1Department of Anaesthesiology, University Medical Centre Regensburg, Regensburg, Germany.

Immunity, Inflammation and Disease
|June 11, 2021
PubMed
Abstract

Insights

Extracellular matrix components significantly alter polymorphonuclear neutrophil (PMN) functions. Type III collagen impacts PMN migration and reactive oxygen species (ROS) production, while other matrices affect myeloperoxidase (MPO) release and NETosis.

Area of Science:

  • Immunology
  • Cell Biology
  • Biomaterials Science

Background:

  • Polymorphonuclear neutrophils (PMN) are crucial for immune responses, including migration, reactive oxygen species (ROS) production, myeloperoxidase (MPO) release, and NETosis.
  • Dysregulated PMN inflammatory responses contribute to disease pathophysiology.
  • The behavior of PMN after migrating through blood vessel walls and their interaction with the extracellular matrix (ECM) remain poorly understood.

Purpose of the Study:

  • To investigate the influence of different extracellular matrix (ECM) components on PMN function.
  • To understand how ECM interactions modulate key PMN immunological reactions.

Main Methods:

  • An in vitro chemotaxis model was established using type I and III collagen, fibrin, and herbal agarose.
  • PMN behavior within these matrices was analyzed using µ-slide chemotaxis devices and N-formylmethionine-leucyl-phenylalanine (fMLP) stimulation.
  • Live-cell imaging with fluorescent time-lapse microscopy was employed to assess PMN function.

Main Results:

  • The ECM significantly influenced PMN functions.
  • Type III collagen inhibited PMN migration (track length, direction, targeting) but accelerated ROS production.
  • Agarose inhibited MPO release, and fibrin retarded NETosis.

Conclusions:

  • The interaction between PMN and ECM components modulates critical immune functions.
  • Given the prevalence of type III collagen in lung and skin, these interactions may play a role in diseases like acute respiratory distress syndrome and pyoderma gangrenosum.
Keywords:
extracellularinflammatory diseasematrixneutrophil functiontissue

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