Reconstruction of the Mouse Inflammasome System in HEK293T Cells

Bio-protocol
Hexin ShiBruce Beutler

Abstract

The NLRP3 (NLR family, Pyrin domain containing 3) inflammasome is a multiprotein complex comprised of NLRP3, pro-caspase-1, the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), and the protein kinase NIMA related kinase 7 (NEK7) (Shi et al., 2016; He et al., 2016; Schmid-Burgk et al., 2016). When cells are exposed to microbes and/or danger signals, the inflammasome assembles and serves as a platform for the activation of caspase-1. Caspase-1 activation promotes the processing and secretion of the pro-inflammatory cytokines interleukin-1β (IL-1β), IL-18, and IL-33 as well as pyroptosis induction (Gross et al., 2011; Arend et al., 2008), which elicit inflammatory responses. Here, we describe how to co-transfect the NLRP3 inflammasome components into HEK293T cells, which enables inflammasome activation and the production of IL-1β upon stimulation with nigericin.

References

Jul 11, 2008·Immunological Reviews·William P ArendCem Gabay
Sep 3, 2011·Immunological Reviews·Olaf GrossJurg Tschopp
Jul 18, 2012·Nature·Ben LuKevin J Tracey

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Citations

Sep 29, 2020·Frontiers in Immunology·Tabassum RahmanJonathan A Harton
Jun 21, 2020·International Journal of Molecular Sciences·Giovanni ZitoChiara Cipollina
Aug 14, 2020·Cell Metabolism·Alexander HooftmanLuke A J O'Neill
Nov 9, 2021·Frontiers in Immunology·Abhinit NagarJonathan A Harton

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis