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Updated: Nov 15, 2025
Isolation and Characterization of Neutrophils with Anti-Tumor Properties
Published on: June 19, 2015
Phenotypical and Functional Characterization of Neutrophils in Two Pyrin-Associated Auto-inflammatory Diseases
Bert Malengier-Devlies1, Mieke Metzemaekers2, Mieke Gouwy2
1Laboratory of Immunobiology, Rega Institute, KU Leuven, Leuven, Belgium.
Purpose:
Familial Mediterranean Fever (FMF) and Pyrin-Associated Autoinflammation with Neutrophilic Dermatosis (PAAND) are clinically distinct autoinflammatory disorders caused by mutations in the pyrin-encoding gene MEFV. We investigated the transcriptional, phenotypical, and functional characteristics of patient neutrophils to explore their potential role in FMF and PAAND pathophysiology.
Methods:
RNA sequencing was performed to discover transcriptional aberrancies. The phenotypical features, degranulation properties, and phagocytic capacity of neutrophils were assessed by flow cytometry. Production of reactive oxygen species (ROS), myeloperoxidase (MPO) release, and chemotactic responses were investigated via chemiluminescence, ELISA, and Boyden chamber assays, respectively.
Results:
Neutrophils from PAAND and FMF patients showed a partially overlapping, activated gene expression profile with increased expression of S100A8, S100A9, S100A12, IL-4R, CD48, F5, MMP9, and NFKB. Increased MMP9 and S100A8/A9 expression levels were accompanied by high plasma concentrations of the encoded proteins. Phenotypical analysis revealed that neutrophils from FMF patients exhibited an immature character with downregulation of chemoattractant receptors CXCR2, C5aR, and BLTR1 and increased expression of Toll-like receptor 4 (TLR4) and TLR9. PAAND neutrophils displayed an increased random, but reduced CXCL8-induced migration. A tendency for enhanced random migration was observed for FMF neutrophils. PAAND neutrophils showed a moderately but significantly enhanced phagocytic activity as opposed to neutrophils from FMF patients. Neutrophils from both patient groups showed increased MPO release and ROS production.
Conclusions:
Neutrophils from patients with FMF and PAAND, carrying different mutations in the MEFV gene, share a pro-inflammatory phenotype yet demonstrate diverse features, underscoring the distinction between both diseases.
Insights
Neutrophils in Familial Mediterranean Fever (FMF) and Pyrin-Associated Autoinflammation with Neutrophilic Dermatosis (PAAND) share inflammatory traits but show distinct characteristics, highlighting differences in these autoinflammatory disorders.
Area of Science:
- Immunology
- Genetics
- Molecular Biology
Background:
- Familial Mediterranean Fever (FMF) and Pyrin-Associated Autoinflammation with Neutrophilic Dermatosis (PAAND) are distinct autoinflammatory diseases.
- Both conditions stem from mutations in the pyrin-encoding gene MEFV.
- Understanding neutrophil behavior is crucial for elucidating disease pathophysiology.
Purpose of the Study:
- To investigate the transcriptional, phenotypical, and functional characteristics of neutrophils in FMF and PAAND patients.
- To explore the role of neutrophils in the distinct pathologies of FMF and PAAND.
Main Methods:
- RNA sequencing to identify transcriptional differences.
- Flow cytometry for phenotypical analysis and degranulation.
- Chemiluminescence, ELISA, and Boyden chamber assays to assess reactive oxygen species (ROS), myeloperoxidase (MPO) release, and chemotaxis.
Main Results:
- Neutrophils from both FMF and PAAND patients displayed a shared pro-inflammatory gene expression profile, including elevated S100A8/A9.
- FMF neutrophils showed immature characteristics with altered receptor expression, while PAAND neutrophils exhibited altered migration and enhanced phagocytosis.
- Both groups demonstrated increased MPO release and ROS production.
Conclusions:
- Neutrophils in FMF and PAAND share a pro-inflammatory phenotype due to MEFV mutations.
- Despite shared features, distinct neutrophil characteristics underscore the clinical differences between FMF and PAAND.
