ATS 2021 will feature presentations of original research from accepted abstracts. Mini Symposia and Thematic Poster Sessions are abstract based sessions.
Please use the form below to browse scientific abstracts and case reports accepted for ATS 2021. Abstracts presented at the ATS 2021 will be published in the Online Abstract Issue of the American Journal of Respiratory and Critical Care Medicine, Volume 203, May 3, 2021.
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Extracellular Specks, a Marker of NLRP3 Inflammasome Activation
Session Title
TP4 - TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES
Abstract
A1318 - Extracellular Specks, a Marker of NLRP3 Inflammasome Activation
Author Block: M. Rhedin1, Z. Jevnikar Rojnik2, B. Collins3, J. Jirholt3; 1Bioscience COPD/IPF, Research and Early Development, Respiratory and Immunology (R&I), BioPharmaceu, AstraZeneca, Gothenburg, Sweden, 2Translational Science and Experimental Medicine, Research and Early Development, Respiratory & Immun, AstraZeneca, Gothenburg, Sweden, 3AstraZeneca, Gothenburg, Sweden.
Introduction: The NLRP3 inflammasome has been implicated in driving pathological damage and chronic inflammation in a number of diseases including COPD. In response to cellular stress the NLRP3 inflammasome condenses into speck structures containing NLRP3, ASC and Caspase-1. Assembly of specks lead to Caspase-1 activation and sequent IL-1β maturation as well as cleavage of gasdermin leading to pyroptosis (cell death). Since it has been demonstrated that these speck structures can be found extracellularly they could possibly be used to detect NLRP3 activation.
Methods: Using cells expressing labeled ASC (THP-1-ASC-GFP, InvivoGen), part of the NLRP3 inflammasome, we evaluated an assay to detect extracellular specks (eSpecks) in vitro. In addition we verified the function of an anti ASC antibody to be able to measure eSpecks from unlabeled cells.
Results: We used flow cytometry to detect eSpecks secreted into the medium of THP-1-ASC-GFP cells, THP-1 cells and human primary monocytes subjected to NLRP3 activation.
Conclusions: We evaluated an commercially available antibody for use in the detection of eSpecks. Further work using patient material is necessary to evaluate the usefulness of this approach to detect activation of NLRP3 inflammasome.
Author Block: M. Rhedin1, Z. Jevnikar Rojnik2, B. Collins3, J. Jirholt3; 1Bioscience COPD/IPF, Research and Early Development, Respiratory and Immunology (R&I), BioPharmaceu, AstraZeneca, Gothenburg, Sweden, 2Translational Science and Experimental Medicine, Research and Early Development, Respiratory & Immun, AstraZeneca, Gothenburg, Sweden, 3AstraZeneca, Gothenburg, Sweden.
Introduction: The NLRP3 inflammasome has been implicated in driving pathological damage and chronic inflammation in a number of diseases including COPD. In response to cellular stress the NLRP3 inflammasome condenses into speck structures containing NLRP3, ASC and Caspase-1. Assembly of specks lead to Caspase-1 activation and sequent IL-1β maturation as well as cleavage of gasdermin leading to pyroptosis (cell death). Since it has been demonstrated that these speck structures can be found extracellularly they could possibly be used to detect NLRP3 activation.
Methods: Using cells expressing labeled ASC (THP-1-ASC-GFP, InvivoGen), part of the NLRP3 inflammasome, we evaluated an assay to detect extracellular specks (eSpecks) in vitro. In addition we verified the function of an anti ASC antibody to be able to measure eSpecks from unlabeled cells.
Results: We used flow cytometry to detect eSpecks secreted into the medium of THP-1-ASC-GFP cells, THP-1 cells and human primary monocytes subjected to NLRP3 activation.
Conclusions: We evaluated an commercially available antibody for use in the detection of eSpecks. Further work using patient material is necessary to evaluate the usefulness of this approach to detect activation of NLRP3 inflammasome.
