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. 2018 Apr 5;73(8):758–768. doi: 10.1136/thoraxjnl-2017-210326

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© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

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Phosphoinositide 3-kinase (PI3K)-δ regulates Aspergillus fumigatus (Af)-induced nucleotide-binding domain, leucine-rich-containing family, pyrin-domain-containing-3 (NLRP3) inflammasome assembly/activation in normal human bronchial epithelial (NHBE) cells. (A and B) Representative confocal images of NHBE cells show the localisation of NLRP3 (red), caspase-1 (Casp-1; green) and apoptosis-associated speck-like protein containing a carboxy-terminal caspase-recruitment domain (ASC; green) in the control (no treatment), Af-stimulated cells (Af), Af-stimulated cells administered drug vehicle (Af+VEH), or Af-stimulated cells administered IC87114 (Af+IC). 4',6-Diamidino-2-phenylindole (DAPI) stain was used for nuclear localisation. Bars indicate 20 µm. (C) Representative immunoblot of ASC monomers and high order complexes in cross-linked cytosolic pellets from Af-stimulated NHBE cells in the presence and absence of IC87114. (D and E) Representative immunoblots and densitometric analyses of NLRP3 (D) and IL-1β (E) after stimulation with Af in NHBE cells in the presence or absence of IC87114. Bars represent mean±SEM from three independent experiments. ^# P<0.05 versus control; *P<0.05 versus cells stimulated with Af alone. (F and G) Representative immunoblots and densitometric analyses of NLRP3 (F) and IL-1β (G) after stimulation with Af in NHBE cells transfected with either non-targeting siRNA (NT siRNA) or PI3K-δ siRNA. Bars represent mean±SEM from three independent experiments. ^# P<0.05 versus control; *P<0.05 versus cells stimulated with Af transfected with NT siRNA. (H) Representative confocal images of NHBE cells show the localisation of NLRP3 (red) and Casp-1 (green) in the control (no treatment), Af-stimulated cells (Af), Af-stimulated cells administered NT siRNA (Af+NT), or Af-stimulated cells administered PI3K-δ siRNA (Af+PI3K-δ). Bars indicate 20 µm. (I) Representative confocal images of NHBE cells show the localisation of mitochondrial reactive oxygen species (mtROS) in the control, Af, Af+VEH or Af+IC. Bars indicate 20 µm.