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Updated: Mar 31, 2026
A High-throughput Assay to Assess and Quantify Neutrophil Extracellular Trap Formation
Published on: January 29, 2019
Dengue Virus Serotype-2 Interferes with the Formation of Neutrophil Extracellular Traps
Maria Maximina B Moreno-Altamirano1, Oscar Rodríguez-Espinosa, Oscar Rojas-Espinosa
1Laboratorio de Inmunorregulacix00F3;n, Departamento de Inmunologx00ED;a, Escuela Nacional de Ciencias Biolx00F3;gicas, Instituto Politx00E9;cnico Nacional, Mexico City, Mexico.
Objectives:
Neutrophils play an important role in the control of pathogens through several mechanisms, including phagocytosis and the formation of neutrophil extracellular traps (NETs). The latter consists of DNA as a backbone with embedded antimicrobial peptides, histones, and proteases, providing a matrix to entrap and in some cases to kill microbes. Some metabolic requirements for NET formation have recently been described. The virus-induced formation of NETs and the role of these traps in viral infections remain scarcely reported. Here, we analyzed whether dengue virus serotype-2 (DENV-2) induces NET formation and the DENV-2 effect on phorbol myristate acetate (PMA)-induced NETs.
Methods:
Peripheral blood-derived neutrophils were exposed in vitro to DENV-2 or exposed to DENV-2 and then stimulated with PMA. NET formation was assessed by fluorescence microscopy. Cell membrane Glut-1, glucose uptake, and reactive oxygen species (ROS) production were assessed.
Results:
DENV-2 does not induce the formation of NETs. Moreover, DENV-2 inhibits PMA-induced formation of NETs by about 80%. This effect is not related to the production of ROS. The mechanism seemingly accountable for this inhibitory effect is the DENV-2-mediated inhibition of PMA-induced glucose uptake by neutrophils.
Conclusion:
Our results suggest that DENV-2 inhibits glucose uptake as a metabolism-based way to avoid the formation of NETs.
Insights
Dengue virus serotype-2 (DENV-2) inhibits neutrophil extracellular trap (NET) formation by blocking glucose uptake. This metabolic interference prevents neutrophils from effectively trapping pathogens during DENV-2 infection.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Neutrophils combat pathogens via phagocytosis and neutrophil extracellular traps (NETs).
- NETs are DNA structures containing antimicrobial components crucial for trapping microbes.
- The role of NETs in viral infections, particularly dengue virus, is not well understood.
Purpose of the Study:
- To investigate if dengue virus serotype-2 (DENV-2) induces NET formation.
- To determine DENV-2's effect on phorbol myristate acetate (PMA)-induced NET formation.
- To elucidate the metabolic mechanisms underlying DENV-2's impact on NETs.
Main Methods:
- In vitro exposure of human neutrophils to DENV-2.
- Assessment of NET formation using fluorescence microscopy.
- Measurement of glucose uptake, Glut-1 expression, and reactive oxygen species (ROS) production.
Main Results:
- DENV-2 does not induce NET formation.
- DENV-2 significantly inhibits PMA-induced NET formation by approximately 80%.
- This inhibition is independent of ROS production but linked to suppressed glucose uptake.
Conclusions:
- DENV-2 actively inhibits NET formation, suggesting a viral immune evasion strategy.
- The mechanism involves DENV-2 interfering with neutrophil glucose metabolism.
- This metabolic disruption hinders the formation of NETs, impacting pathogen control.
