Whole blood assay as a model for in vitro evaluation of inflammasome activation and subsequent caspase-mediated interleukin-1 beta release.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
26
06
2018
accepted:
26
03
2019
entrez:
9
4
2019
pubmed:
9
4
2019
medline:
24
12
2019
Statut:
epublish
Résumé
Processing of pro-interleukin (IL)-1β and IL-18 is regulated by multiprotein complexes, known as inflammasomes. Inflammasome activation results in generation of bioactive IL-1β and IL-18, which can exert potent pro-inflammatory effects. Our aim was to develop a whole blood-based assay to study the inflammasome in vitro and that also can be used as an assay in clinical studies. We show whole blood is a suitable milieu to study inflammasome activation in primary human monocytes. We demonstrated that unprocessed human blood cells can be stimulated to activate the inflammasome by the addition of adenosine 5'-triphosphate (ATP) within a narrow timeframe following lipopolysaccharide (LPS) priming. Stimulation with LPS resulted in IL-1β release; however, addition of ATP is necessary for "full-blown" inflammasome stimulation resulting in high IL-1β and IL-18 release. Intracellular cytokine staining demonstrated monocytes are the major producers of IL-1β in human whole blood cultures, and this was associated with activation of caspase-1/4/5, as detected by a fluorescently labelled caspase-1/4/5 probe. By applying caspase inhibitors, we show that both the canonical inflammasome pathway (via caspase-1) as well as the non-canonical inflammasome pathway (via caspases-4 and 5) can be studied using this whole blood-based model.
Identifiants
pubmed: 30958862
doi: 10.1371/journal.pone.0214999
pii: PONE-D-18-18667
pmc: PMC6453527
doi:
Substances chimiques
IL1B protein, human
0
Inflammasomes
0
Interleukin-18
0
Interleukin-1beta
0
Lipopolysaccharides
0
Caspases
EC 3.4.22.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0214999Déclaration de conflit d'intérêts
Good Biomarker Sciences provided salaries for authors [TATT, KL, KEM, DT]. This commercial affiliation does not alter our adherence to PLOS ONE polices on sharing data and materials.
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