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Quantifying neutrophil extracellular trap release in a combined infection-inflammation NET-array device.

Udaya Sree Datla1,2,3, Bhaskar Vundurthy4, Jessica S Hook5

  • 1Translational Biology, Medicine and Health, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.

Lab on a Chip
|January 8, 2024
PubMed
Summary
This summary is machine-generated.

A new device quantifies neutrophil extracellular traps (NETs) at single-cell resolution. This tool reveals how infection and inflammation influence NET release in different microenvironments, aiding immunotherapy development.

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Area of Science:

  • Immunology
  • Biomedical Engineering
  • Cell Biology

Background:

  • Neutrophil extracellular traps (NETs) are implicated in pathologies like COVID-19, with elevated levels indicating coagulopathy and immunothrombosis risks.
  • Current methods for NET quantification use bulk measurements in simplified settings, lacking single-cell resolution and microenvironment context.
  • Understanding NET release dynamics is crucial for developing targeted immunotherapies.

Purpose of the Study:

  • To develop and validate a novel NET-array device for single-cell resolution quantification of NET release.
  • To investigate the impact of infection (Pseudomonas aeruginosa) and inflammatory mediators on NET release in varied microenvironments.
  • To analyze the spatiotemporal dynamics and characteristics of NET release.

Main Methods:

  • Fabrication of a microfluidic NET-array device with wide chambers and constricted loops.
  • Culture of primary human neutrophils with Pseudomonas aeruginosa PAO1 and inflammatory mediators (TNF-α, IL-6, LTB4).
  • Time-lapse imaging and computer-vision-based image processing for automated quantification of individual NETs and their characteristics.

Main Results:

  • The NET-array device enabled single-cell resolution imaging and quantification of NET release.
  • Combined infection with Pseudomonas aeruginosa and inflammatory mediators (TNF-α, IL-6) significantly increased NET release compared to infection alone.
  • NET release was reduced in constricted microenvironments within the device, and variable NET sizes were observed.
  • Temporal dynamics and relative areas of NETs were quantified, revealing distinct release patterns under different conditions.

Conclusions:

  • The novel NET-array device provides a powerful tool for studying NET release dynamics at single-cell resolution in defined microenvironments.
  • This system enhances our understanding of how infection and inflammation interact to modulate NETosis.
  • The device holds potential for high-throughput screening of immunotherapies targeting neutrophil-driven inflammation.