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Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
Published on: May 22, 2014
Increased NLRP3-dependent interleukin 1β secretion in patients with familial Mediterranean fever: correlation with
Alessia Omenetti1, Sonia Carta, Laura Delfino
1Pediatrics II Unit, G Gaslini IRCCS and University of Genoa, , Genoa, Italy.
Objectives:
To define in patients affected by familial Mediterranean fever (FMF) whether or not interleukin (IL)-1β secretion (1) is enhanced, (2) correlates with the type of MEFV mutation and (3) is mediated by NLRP3.
Methods:
Freshly isolated monocytes from 21 patients with FMF (12 homozygous and 9 heterozygous), 14 MEFV healthy carriers and 30 healthy donors (HDs), unstimulated or after lipopolysaccharide (LPS)-induced activation, were analysed for redox state (production of reactive oxygen species (ROS) and antioxidant responses) and IL-1β and IL-1 receptor antagonist (IL-1Ra) secretion. NLRP3 down-modulation was induced by in vitro silencing of the NLRP3 gene.
Results:
LPS-stimulated monocytes from patients with FMF displayed enhanced IL-1β secretion, which correlated with number and penetrance of MEFV mutations. Silencing of NLRP3 consistently inhibited IL-1β secretion. As in other autoinflammatory diseases, FMF monocytes produced more ROS than genetically negative cells from HDs. Unlike in cryopyrin-associated periodic fever syndromes (CAPS), however, they were characterised by a conserved and sustained antioxidant response. Consistent with this finding, activated MEFV-mutated monocytes did not exhibit the functional indicators of oxidative stress observed in CAPS, including accelerated IL-1β secretion and deficient production of IL-1Ra.
Conclusions:
MEFV-mutated monocytes display enhanced IL-1β secretion, which correlates with number of high-penetrance mutations and level of endogenous ROS. Unlike NLRP3-mutated cells, monocytes carrying MEFV mutations withstand oxidative stress and preserve IL-1Ra production, thereby limiting inflammation. Finally, in contrast with that found in the animal model, the increased secretion of IL-1β by LPS-stimulated FMF monocytes is NLRP3-dependent.
Insights
Familial Mediterranean Fever (FMF) patients show increased interleukin-1β secretion, linked to MEFV mutations and NLRP3 inflammasome activation. FMF monocytes resist oxidative stress, preserving anti-inflammatory responses.
Area of Science:
- Immunology
- Genetics
- Molecular Biology
Background:
- Familial Mediterranean Fever (FMF) is a genetic autoinflammatory disorder characterized by recurrent inflammatory episodes.
- The MEFV gene mutations are responsible for FMF, leading to dysregulation of inflammatory pathways.
- Interleukin-1β (IL-1β) is a key pro-inflammatory cytokine implicated in FMF pathogenesis.
Purpose of the Study:
- To investigate IL-1β secretion in FMF patients.
- To determine the correlation between MEFV mutation type and IL-1β secretion.
- To elucidate the role of NLRP3 inflammasome in IL-1β production in FMF.
Main Methods:
- Monocytes from FMF patients, healthy carriers, and healthy donors were analyzed.
- Lipopolysaccharide (LPS) was used to stimulate monocytes.
- Reactive oxygen species (ROS) production, antioxidant responses, IL-1β, and IL-1 receptor antagonist (IL-1Ra) secretion were measured.
- NLRP3 gene silencing was performed to assess its role.
Main Results:
- LPS-stimulated FMF monocytes exhibited enhanced IL-1β secretion, correlating with MEFV mutation load and penetrance.
- NLRP3 silencing significantly reduced IL-1β secretion, confirming its involvement.
- FMF monocytes showed increased ROS production but maintained antioxidant responses, unlike in cryopyrin-associated periodic fever syndromes (CAPS).
- MEFV-mutated monocytes preserved IL-1Ra production despite activation.
Conclusions:
- MEFV-mutated monocytes demonstrate heightened IL-1β secretion, dependent on NLRP3 and correlated with mutation severity and ROS levels.
- FMF monocytes possess a unique ability to withstand oxidative stress and maintain IL-1Ra production, contributing to inflammation control.
- The increased IL-1β secretion in FMF is NLRP3-dependent, contrasting with some animal models.
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