Establishing a J774A.1 Murine Macrophage NLRP3 Inflammasome Model for Examining Short-Chain Fatty Acid Suppression

Authors

  • Samantha N. Cortez Department of Microbiology and Immunology, University of British Columbia
  • Jerry Y. He Undergraduate Research Opportunities (URO)
  • Vince M. Lising Department of Microbiology and Immunology, University of British Columbia

Abstract

The NLRP3 inflammasome is an assembly of effector and adaptor proteins that mediates the release of pro-inflammatory cytokines, making it a key component of inflammation. While the inflammasome plays essential roles in pathogen clearance and homeostasis, it has also been linked to inflammatory diseases such as diabetes. With the increasing prevalence of obesity and type 2 diabetes, the inflammasome acts as a promising target for potential therapeutics. Recent studies have pointed to the role short-chain fatty acids might have in suppressing inflammation and the inflammasome. In this study, we set out to confirm whether the short-chain fatty acid, butyrate, has a suppressive effect on the NLRP3 inflammasome. We observed priming of the inflammasome in LPS-primed, ATP-activated J774A.1 macrophages through increased pro-IL-1β. However, mature IL-1β after ATP stimulation was not detected in our western blot analyses. Our cell viability and LDH cytotoxicity assays confirm ATP activation of the inflammasome through pyroptosis and LDH release. The establishment of a working NLRP3 model for J774A.1 macrophages will allow for research on the suppressive effects of short-chain fatty acids on the inflammasome and provide new avenues for potential therapeutics for inflammatory diseases such as diabetes. 

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Published

2022-08-31

Issue

Vol. 27 (2022): Undergraduate Journal of Experimental Microbiology and Immunology (UJEMI)

Section

Research Article