Bert Malengier-Devlies1, Mieke Metzemaekers2, Mieke Gouwy2, Erika Van Nieuwenhove3, Albrecht Betrains4,5, Maaike Cockx2, Lotte Vanbrabant2, Noëmie Pörtner2, Jurgen Vercauteren6, Lien De Somer1,3,5, Sofie Struyf2, Steven Vanderschueren4,5,7, Ellen De Langhe5,8,9, Paul Proost10, Patrick Matthys1, Carine Wouters11,12,13. 1. Laboratory of Immunobiology, Rega Institute, KU Leuven, Leuven, Belgium. 2. Laboratory of Molecular Immunology, Rega Institute, KU Leuven, Leuven, Belgium. 3. Division of Pediatric Rheumatology, Department Pediatrics, University Hospitals Leuven, Leuven, Belgium. 4. Laboratory of Clinical Infectious and Inflammatory Disorders, KU Leuven, Leuven, Belgium. 5. European Reference Network for Rare Immunodeficiency, Autoinflammatory and Autoimmune Diseases (RITA), University Hospital Leuven, Leuven, Belgium. 6. Laboratory of Clinical en Epidemiological Virology, KU Leuven, Leuven, Belgium. 7. Division of General Internal Medicine, University Hospitals Leuven, Leuven, Belgium. 8. Division of Rheumatology, University Hospitals Leuven, Leuven, Belgium. 9. Laboratory of Tissue Homeostasis and Disease, KU Leuven, Leuven, Belgium. 10. Laboratory of Molecular Immunology, Rega Institute, KU Leuven, Leuven, Belgium. paul.proost@kuleuven.be. 11. Laboratory of Immunobiology, Rega Institute, KU Leuven, Leuven, Belgium. carine.wouters@uzleuven.be. 12. Division of Pediatric Rheumatology, Department Pediatrics, University Hospitals Leuven, Leuven, Belgium. carine.wouters@uzleuven.be. 13. European Reference Network for Rare Immunodeficiency, Autoinflammatory and Autoimmune Diseases (RITA), University Hospital Leuven, Leuven, Belgium. carine.wouters@uzleuven.be.
Abstract
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.
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.
Authors: M Centola; G Wood; D M Frucht; J Galon; M Aringer; C Farrell; D W Kingma; M E Horwitz; E Mansfield; S M Holland; J J O'Shea; H F Rosenberg; H L Malech; D L Kastner Journal: Blood Date: 2000-05-15 Impact factor: 22.113