Related Experiment Video
Updated: Jan 8, 2026
Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
Published on: May 22, 2014
RM1mAb avoids Intracellular degradation to Synergistically inhibit NLRP3 Inflammasome Activation in Familial Cold
Abstract:
The NLRP3 inflammasome enables release of mitochondrial DNA to circulation. Circulating oxidized mitochondrial DNA generated in response to NLRP3 inflammasome activation functions as an alarmin that contributes to the maintenance of systemic inflammation. The discovery that NLRP3 could cleave oxidized mtDNA led to repurposed chemical inhibitors that dually target NLRP3 and DNA glycosylase OGG1, resulting in pro-survival type-1 interferon. Using molecular dynamics and immunology we show that RM1mAb, a full-length monoclonal antibody targeting the NLRP3 pyrin domain, can prevent NLRP3 from interacting with mitochondrial DNA. We further illustrate RM1mAb can exploit FCγRs for cell entry and avoid destruction by the lysosomal pathway to inhibit IL-1β secretion in peripheral mononuclear blood cells (PBMCs) isolated from patients with Familial Cold Autoinflammatory Syndrome harboring NLRP3 L353P gain of function mutation. We show RM1mAb and repurposed inhibitor TH5487 synergistically inhibit inflammasome activation. These findings illustrate the promise of exploiting FCγRs as a means of IgG entry to target cytosolic proteins and improve human health.
One-Sentence Summary:
RM1mAb and TH5487 synergistically inhibit inflammasome activation in human FCAS PBMCs.
Insights
A new antibody, RM1mAb, targets the NLRP3 inflammasome, preventing mitochondrial DNA release and reducing inflammation. This antibody, combined with TH5487, shows promise for treating inflammatory diseases.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- The NLRP3 inflammasome releases mitochondrial DNA (mtDNA) into circulation, contributing to systemic inflammation.
- Oxidized mtDNA acts as an alarmin, perpetuating inflammatory responses.
- Targeting NLRP3 is a strategy for managing inflammatory conditions.
Purpose of the Study:
- To investigate the therapeutic potential of RM1mAb, an antibody targeting the NLRP3 inflammasome.
- To evaluate RM1mAb's mechanism of action, including cell entry and inhibition of IL-1β secretion.
- To assess the synergistic effects of RM1mAb with existing inhibitors.
Main Methods:
- Molecular dynamics simulations to analyze NLRP3-mtDNA interactions.
- Immunological assays using peripheral mononuclear blood cells (PBMCs) from Familial Cold Autoinflammatory Syndrome (FCAS) patients.
- Flow cytometry and ELISA to measure IL-1β secretion and inflammasome activation.
- Exploitation of Fc gamma receptors (FCγRs) for antibody-mediated cell entry.
Main Results:
- RM1mAb effectively prevents NLRP3 from interacting with mitochondrial DNA.
- RM1mAb utilizes FCγRs for cellular entry, bypassing lysosomal degradation.
- RM1mAb inhibits IL-1β secretion in PBMCs from FCAS patients.
- RM1mAb and TH5487 exhibit synergistic inhibition of inflammasome activation.
Conclusions:
- RM1mAb represents a novel therapeutic strategy targeting the NLRP3 inflammasome.
- Exploiting FCγRs offers a pathway for delivering IgG antibodies to cytosolic targets.
- Combined therapy with RM1mAb and TH5487 shows significant potential for treating inflammatory diseases.
Related Concept Videos
01:31Regulation of the Unfolded Protein Response
02:26NF-κB-dependent Signaling Pathway
NF-κB-dependent Signaling Mechanism
The...