PMID- 23852597
OWN - NLM
STAT- MEDLINE
DCOM- 20140207
LR  - 20171116
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1040
DP  - 2013
TI  - Measuring inflammasome activation in response to bacterial infection.
PG  - 65-84
LID - 10.1007/978-1-62703-523-1_6 [doi]
AB  - Inflammasomes are multi-protein signaling platforms assembled in response to 
      viral and bacterial pathogens as well as endogenous danger signals. Inflammasomes 
      serve as activation platforms for the mammalian cysteine protease caspase-1, a 
      central mediator of innate immunity. The hallmarks of inflammasome activation are 
      the processing of caspase-1, the maturation and release of interleukin-1beta (IL-1beta) 
      and the induction of pyroptosis, a lytic inflammatory cell death. This protocol 
      describes methods for studying inflammasome activation in response to bacterial 
      pathogens in bone-marrow derived murine macrophages (BMDMs). In particular, we 
      outline the protocols to measure cytokine maturation by ELISA and pyroptosis by 
      the release of Lactate Dehydrogenase (LDH). In addition, we describe methods to 
      visualize endogenous ASC specks or foci in infected cells and to study the 
      release of processed caspase-1, caspase-11 and mature cytokines into the cell 
      supernatant by Western blotting. General considerations are discussed to design 
      and optimize the infection protocol for the study of inflammasome activation by 
      other bacterial pathogens.
FAU - Broz, Petr
AU  - Broz P
AD  - Department of Microbiology and Immunology, Stanford School of Medicine, Stanford 
      University, Stanford, CA, USA.
FAU - Monack, Denise M
AU  - Monack DM
LA  - eng
GR  - AI08972/AI/NIAID NIH HHS/United States
GR  - AI095396/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Apoptosis Regulatory Proteins)
RN  - 0 (CARD Signaling Adaptor Proteins)
RN  - 0 (Cytokines)
RN  - 0 (Cytoskeletal Proteins)
RN  - 0 (Inflammasomes)
RN  - 0 (Pycard protein, mouse)
RN  - EC 3.4.22.36 (Caspase 1)
SB  - IM
MH  - Animals
MH  - Apoptosis Regulatory Proteins
MH  - Bacterial Infections/immunology/*metabolism
MH  - Blotting, Western
MH  - CARD Signaling Adaptor Proteins
MH  - Caspase 1/metabolism
MH  - Cell Differentiation
MH  - Cytokines/metabolism
MH  - Cytoskeletal Proteins/metabolism
MH  - Inflammasomes/*metabolism
MH  - Macrophages/cytology/immunology/metabolism/microbiology
MH  - Mice
MH  - Microscopy, Confocal
EDAT- 2013/07/16 06:00
MHDA- 2014/02/08 06:00
CRDT- 2013/07/16 06:00
PHST- 2013/07/16 06:00 [entrez]
PHST- 2013/07/16 06:00 [pubmed]
PHST- 2014/02/08 06:00 [medline]
AID - 10.1007/978-1-62703-523-1_6 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2013;1040:65-84. doi: 10.1007/978-1-62703-523-1_6.