Exposing kinetic disparities between inflammasome readouts using time-resolved analysis
Matthew Herring,
Alexander Persson,
Ryan Potter,
Roger Karlsson,
Eva Särndahl,
Mikael Ejdebäck
Affiliations
Matthew Herring
School of Medical Sciences, Faculty of Medicine and Health, Örebro University, Örebro, Sweden; Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, Örebro, Sweden; School of Bioscience, Systems Biology Research Centre, University of Skövde, Skövde, Sweden; Corresponding author. School of Medical Sciences, Faculty of Medicine and Health, Örebro University, Örebro, Sweden
Alexander Persson
School of Medical Sciences, Faculty of Medicine and Health, Örebro University, Örebro, Sweden; Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, Örebro, Sweden
Ryan Potter
School of Bioscience, Systems Biology Research Centre, University of Skövde, Skövde, Sweden; Department of Clinical Neuroscience, Institute of Neuroscience and Physiology, Sahlgrenska Academy, Göteborg University, Göteborg, Sweden
Roger Karlsson
Nanoxis Consulting AB, Göteborg, Sweden; Department of Infectious Diseases, Institute of Biomedicine, Sahlgrenska Academy, Göteborg University, Göteborg, Sweden; Department of Clinical Microbiology, Sahlgrenska University Hospital, Region Västra Götaland, Göteborg, Sweden
Eva Särndahl
School of Medical Sciences, Faculty of Medicine and Health, Örebro University, Örebro, Sweden; Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, Örebro, Sweden
Mikael Ejdebäck
School of Bioscience, Systems Biology Research Centre, University of Skövde, Skövde, Sweden
The NLRP3 inflammasome is an intracellular multiprotein complex described to be involved in both an effective host response to infectious agents and various diseases. Investigation into the NLRP3 inflammasome has been extensive in the past two decades, and often revolves around the analysis of a few specific readouts, including ASC-speck formation, caspase-1 cleavage or activation, and cleavage and release of IL-1β and/or IL-18. Quantification of these readouts is commonly undertaken as an endpoint analysis, where the presence of each positive outcome is assessed independently of the others. In this study, we apply time-resolved analysis of a human macrophage model (differentiated THP-1-ASC-GFP cells) to commonly accessible methods. This approach yields the additional quantifiable metrics time-resolved absolute change and acceleration, allowing comparisons between readouts. Using this methodological approach, we reveal (potential) discrepancies between inflammasome-related readouts that otherwise might go undiscovered. The study highlights the importance of time-resolved data in general and may be further extended as well as incorporated into other areas of research.
