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Understanding the Role of the Inflammasome in Inflammation

This webinar will discuss:

  • Two important outcomes of inflammasome induction, caspase-1 induction and pyroptosis
  • A novel method for directly measuring caspase-1 activation

 

Summary

The inflammasome is a multiprotein complex expressed in myeloid cells responsible for activation of inflammatory processes as part of the innate immune system. The inflammasome complex contains different proteins depending on the initiating event, but central to the inflammatory process is activation of caspase-1, which in turn activates the cytokines interleukin-1ß and interleukin-18 by proteolytic cleavage of pro-forms of these molecules. Caspase-1 activation also induces pyroptosis, an inflammatory form of cell death. Typically, inflammasome activation is directly monitored with western blotting of processed caspase-1 or indirectly via determination of IL-1b or IL-18 processing or release, using Western blots or ELISAs, respectively. A bioluminescent assay for the rapid detection of caspase-1 activation has been developed that enables direct measurement of an upstream signaling event in the process of inflammation. We will discuss in detail the specificity of this novel assay, monitoring caspase-1 in cells and released into the medium, differences between cell types, and multiplexing with other assays to measure relevant activities, such as cell death and IL-1b release. This new approach enables studies on the kinetics of inflammasome activation and inactivation, allows a more precise assessment of caspase-1 mediated pyroptosis, and provides a means for high throughput screening of inflammasome modulators.


Speaker

martha-obrien-125x125

Martha OBrien, PhD
Senior Research Scientist
Promega Corporation

Martha O’Brien is a Senior Research Scientist in the Assay Design Group at Promega and is currently leading assay development efforts in the area of inflammation. Martha received her PhD in biology from the University of North Carolina. Prior to joining Promega, she was a postdoctoral fellow and research associate in the Anatomy and Neurobiology Department at Washington University School of Medicine investigating the genetics of neuropeptide systems in Drosophila. As a Promega R&D scientist, she turned to mammalian systems and has developed numerous products, focusing on apoptosis and protease assays, has co-authored several articles and book chapters, and is a co-inventor on several patents.

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