PMID- 37578723
OWN - NLM
STAT- MEDLINE
DCOM- 20230815
LR  - 20230818
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2696
DP  - 2023
TI  - Methods to Activate the NLRP3 Inflammasome.
PG  - 169-197
LID - 10.1007/978-1-0716-3350-2_12 [doi]
AB  - The inflammasome-nucleating cytoplasmic sensor protein NLRP3 (NACHT-, LRR, and 
      PYD domains-containing protein 3, also known as NOD-like receptor pyrin 
      domain-containing 3, NALP3, or cryopyrin) is triggered by a broad spectrum of 
      sterile endogenous danger signals and environmental irritants. Upon activation, 
      NLRP3 engages the adapter protein ASC that in turn recruits the third 
      inflammasome component, the protease caspase-1. Subsequent caspase-1 activation 
      leads to its auto-processing and maturation of the leaderless IL-1 family 
      cytokines IL-1beta and IL-18 as well as cleavage of the pore-forming protein 
      Gasdermin D (GSDMD). GSDMD plasma membrane pores, formed by its N-terminus, 
      facilitate IL-1 release and, typically, subsequent cell lysis (pyroptosis). This 
      protocol explains standard methods, which are routinely used in our laboratory to 
      study NLRP3 inflammasome biology in vitro. It includes experimental approaches 
      using primary murine bone marrow-derived macrophages (BMDMs) and bone 
      marrow-derived dendritic cells (BMDCs), human peripheral blood mononuclear cells 
      (PBMCs), as well as inflammasome-competent cell lines (HoxB8 and THP-1 cells). 
      The protocol covers the use of a broad spectrum of established NLRP3 activators 
      and outlines the use of common inhibitors blocking NLRP3 itself or its upstream 
      triggering events. We also provide guidelines for experimental set-up and crucial 
      experimental controls to investigate NLRP3 inflammasome signaling or study new 
      activators and inhibitors.
CI  - (c) 2023. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Saller, Benedikt S
AU  - Saller BS
AD  - Faculty of Medicine, Institute of Neuropathology, Medical Center - University of 
      Freiburg, Freiburg, Germany.
AD  - Faculty of Biology, University of Freiburg, Freiburg, Germany.
FAU - Neuwirt, Emilia
AU  - Neuwirt E
AD  - Faculty of Medicine, Institute of Neuropathology, Medical Center - University of 
      Freiburg, Freiburg, Germany.
AD  - Faculty of Biology, University of Freiburg, Freiburg, Germany.
AD  - Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, 
      Germany.
FAU - Gross, Olaf
AU  - Gross O
AD  - Faculty of Medicine, Institute of Neuropathology, Medical Center - University of 
      Freiburg, Freiburg, Germany. olaf.gross@uniklinik-freiburg.de.
AD  - Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, 
      Germany. olaf.gross@uniklinik-freiburg.de.
AD  - Faculty of Medicine, Center for Basics in NeuroModulation (NeuroModulBasics), 
      University of Freiburg, Freiburg, Germany. olaf.gross@uniklinik-freiburg.de.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Inflammasomes)
RN  - 0 (NLR Family, Pyrin Domain-Containing 3 Protein)
RN  - EC 3.4.22.36 (Caspase 1)
RN  - 0 (Interleukin-1)
RN  - 0 (Interleukin-1beta)
SB  - IM
MH  - Mice
MH  - Humans
MH  - Animals
MH  - *Inflammasomes/metabolism
MH  - *NLR Family, Pyrin Domain-Containing 3 Protein/metabolism
MH  - Leukocytes, Mononuclear/metabolism
MH  - Caspase 1/metabolism
MH  - Interleukin-1
MH  - Interleukin-1beta/metabolism
OTO - NOTNLM
OT  - Bone marrow-derived macrophages and dendritic cells
OT  - HoxB8
OT  - Inflammasome
OT  - NLRP3 activators and inhibitors
OT  - Peripheral blood mononuclear cells
OT  - Priming
OT  - THP-1
EDAT- 2023/08/14 12:43
MHDA- 2023/08/15 06:42
CRDT- 2023/08/14 11:32
PHST- 2023/08/15 06:42 [medline]
PHST- 2023/08/14 12:43 [pubmed]
PHST- 2023/08/14 11:32 [entrez]
AID - 10.1007/978-1-0716-3350-2_12 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2023;2696:169-197. doi: 10.1007/978-1-0716-3350-2_12.