title: Silica-induced NLRP3 inflammasome activation in vitro and in rat lungs.
abstract: BACKGROUND
  Mineral particles in the lung cause inflammation and silicosis. In myeloid and bronchial epithelial cells the inflammasome plays a role in responses to crystalline silica. Thioredoxin (TRX) and its inhibitory protein TRX-interacting protein link oxidative stress with inflammasome activation. We investigated inflammasome activation by crystalline silica polymorphs and modulation by TRX in vitro, as well as its localization and the importance of silica surface reactivity in rats.
  
  
  METHODS
  We exposed bronchial epithelial cells and differentiated macrophages to silica polymorphs quartz and cristobalite and measured caspase-1 activity as well as the release of IL-1β, bFGF and HMGB1; including after TRX overexpression or treatment with recombinant TRX. Rats were intratracheally instilled with vehicle control, Dörentruper quartz (DQ12) or DQ12 coated with polyvinylpyridine N-oxide. At days 3, 7, 28, 90, 180 and 360 five animals per treatment group were sacrificed. Hallmarks of silicosis were assessed with Haematoxylin-eosin and Sirius Red stainings. Caspase-1 activity in the bronchoalveolar lavage and caspase-1 and IL-1β localization in lung tissue were determined using Western blot and immunohistochemistry (IHC).
  
  
  RESULTS
  Silica polymorphs triggered secretion of IL-1β, bFGF and HMGB1 in a surface reactivity dependent manner. Inflammasome readouts linked with caspase-1 enzymatic activity were attenuated by TRX overexpression or treatment. At day 3 and 7 increased caspase-1 activity was detected in BALF of the DQ12 group and increased levels of caspase-1 and IL-1β were observed with IHC in the DQ12 group compared to controls. DQ12 exposure revealed silicotic nodules at 180 and 360 days. Particle surface modification markedly attenuated the grade of inflammation and lymphocyte influx and attenuated the level of inflammasome activation, indicating that the development of silicosis and inflammasome activation is determined by crystalline silica surface reactivity.
  
  
  CONCLUSIONS
  Our novel data indicate the pivotal role of surface reactivity of crystalline silica to activate the inflammasome in cultures of both epithelial cells and macrophages. Inhibitory capacity of the antioxidant TRX to inflammasome activation was evidenced. DQ12 quartz exposure induced acute and chronic functional activation of the inflammasome in the heterogeneous cell populations of the lung in associated with its crystalline surface reactivity.
authors.affiliation: Department of Respiratory Medicine, Maastricht University Medical Centre+ (MUMC+), Maastricht University, Maastricht, The Netherlands. p.peeters@maastrichtuniversity.nl.|IUF - Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany. p.peeters@maastrichtuniversity.nl.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Peeters
authors.affiliation: Department of Respiratory Medicine, Maastricht University Medical Centre+ (MUMC+), Maastricht University, Maastricht, The Netherlands. irene.eurlings@maastrichtuniversity.nl.
authors.forename: Irene M J
authors.initials: IM
authors.lastname: Eurlings
authors.affiliation: Department of Respiratory Medicine, Maastricht University Medical Centre+ (MUMC+), Maastricht University, Maastricht, The Netherlands. t.perkins@maastrichtuniversity.nl.
authors.forename: Timothy N
authors.initials: TN
authors.lastname: Perkins
authors.affiliation: Department of Respiratory Medicine, Maastricht University Medical Centre+ (MUMC+), Maastricht University, Maastricht, The Netherlands. e.wouters@mumc.nl.
authors.forename: Emiel F
authors.initials: EF
authors.lastname: Wouters
authors.affiliation: IUF - Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany. Roel.Schins@uni-duesseldorf.de.
authors.forename: Roel P F
authors.initials: RP
authors.lastname: Schins
authors.affiliation: Bèta Sciences and Technology, Hogeschool Zuyd, Heerlen, The Netherlands. paul.borm@zuyd.nl.
authors.forename: Paul J A
authors.initials: PJ
authors.lastname: Borm
authors.affiliation: Histo-Path GmbH, Hannover, Germany. wolfgang.drommer@online.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Drommer
authors.affiliation: Department of Respiratory Medicine, Maastricht University Medical Centre+ (MUMC+), Maastricht University, Maastricht, The Netherlands. n.reynaert@maastrichtuniversity.nl.
authors.forename: Niki L
authors.initials: NL
authors.lastname: Reynaert
authors.affiliation: IUF - Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany. Catrin.Albrecht@IUF-Duesseldorf.de.
authors.forename: Catrin
authors.initials: C
authors.lastname: Albrecht
mesh_terms: D000393:Air Pollutants / Q000737:chemistry / Q000633:toxicity*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001980:Bronchi / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D002352:Carrier Proteins / Q000819:agonists* / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D006801:Humans; D058847:Inflammasomes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D019570:Inhalation Exposure / Q000009:adverse effects; D008168:Lung / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D010316:Particle Size; D051381:Rats; D017208:Rats, Wistar; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D012822:Silicon Dioxide / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity*; D012829:Silicosis / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D013499:Surface Properties; D023382:Toxicity Tests, Acute; D036401:Toxicity Tests, Chronic
chemical_list: D000393:Air Pollutants; D015415:Biomarkers; D002352:Carrier Proteins; D058847:Inflammasomes; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; C573346:Nlrp3 protein, rat; D012822:Silicon Dioxide