title: An Allergic Lung Microenvironment Suppresses Carbon Nanotube-Induced Inflammasome Activation via STAT6-Dependent Inhibition of Caspase-1.
abstract: BACKGROUND
  Multi-walled carbon nanotubes (MWCNTs) represent a human health risk as mice exposed by inhalation display pulmonary fibrosis. Production of IL-1β via inflammasome activation is a mechanism of MWCNT-induced acute inflammation and has been implicated in chronic fibrogenesis. Mice sensitized to allergens have elevated T-helper 2 (Th2) cytokines, IL-4 and IL-13, and are susceptible to MWCNT-induced airway fibrosis. We postulated that Th2 cytokines would modulate MWCNT-induced inflammasome activation and IL-1β release in vitro and in vivo during allergic inflammation.
  
  
  METHODS
  THP-1 macrophages were primed with LPS, exposed to MWCNTs and/or IL-4 or IL-13 for 24 hours, and analyzed for indicators of inflammasome activation. C57BL6 mice were sensitized to house dust mite (HDM) allergen and MWCNTs were delivered to the lungs by oropharyngeal aspiration. Mice were euthanized 1 or 21 days post-MWCNT exposure and evaluated for lung inflammasome components and allergic inflammatory responses.
  
  
  RESULTS
  Priming of THP-1 macrophages with LPS increased pro-IL-1β and subsequent exposure to MWCNTs induced IL-1β secretion. IL-4 or IL-13 decreased MWCNT-induced IL-1β secretion by THP-1 cells and reduced pro-caspase-1 but not pro-IL-1β. Treatment of THP-1 cells with STAT6 inhibitors, either Leflunomide or JAK I inhibitor, blocked suppression of caspase activity by IL-4 and IL-13. In vivo, MWCNTs alone caused neutrophilic infiltration into the lungs of mice 1 day post-exposure and increased IL-1β in bronchoalveolar lavage fluid (BALF) and pro-caspase-1 immuno-staining in macrophages and airway epithelium. HDM sensitization alone caused eosinophilic inflammation with increased IL-13. MWCNT exposure after HDM sensitization increased total cell numbers in BALF, but decreased numbers of neutrophils and IL-1β in BALF as well as reduced pro-caspase-1 in lung tissue. Despite reduced IL-1β mice exposed to MWCNTs after HDM developed more severe airway fibrosis by 21 days and had increased pro-fibrogenic cytokine mRNAs.
  
  
  CONCLUSIONS
  These data indicate that Th2 cytokines suppress MWCNT-induced inflammasome activation via STAT6-dependent down-regulation of pro-caspase-1 and suggest that suppression of inflammasome activation and IL-1β by an allergic lung microenvironment is a mechanism through which MWCNTs exacerbate allergen-induced airway fibrosis.
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Shipkowski
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Alexia J
authors.initials: AJ
authors.lastname: Taylor
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Thompson
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Ellen E
authors.initials: EE
authors.lastname: Glista-Baker
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Brian C
authors.initials: BC
authors.lastname: Sayers
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: Zachary J
authors.initials: ZJ
authors.lastname: Messenger
authors.affiliation: Center for Environmental Medicine, Asthma, and Lung Biology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina, United States of America.
authors.forename: Rebecca N
authors.initials: RN
authors.lastname: Bauer
authors.affiliation: Center for Environmental Medicine, Asthma, and Lung Biology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina, United States of America.
authors.forename: Ilona
authors.initials: I
authors.lastname: Jaspers
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology Program, North Carolina State University, Raleigh, North Carolina, United States of America.
authors.forename: James C
authors.initials: JC
authors.lastname: Bonner
mesh_terms: D000818:Animals; D039741:Antigens, Dermatophagoides / Q000276:immunology; D020170:Caspase 1 / Q000378:metabolism*; D002460:Cell Line; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D005355:Fibrosis; D015870:Gene Expression; D006801:Humans; D006967:Hypersensitivity / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D058847:Inflammasomes / Q000378:metabolism*; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D007958:Leukocyte Count; D008070:Lipopolysaccharides / Q000276:immunology; D008168:Lung / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D037742:Nanotubes, Carbon / Q000009:adverse effects*; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism; D039981:Pyroglyphidae / Q000276:immunology; D050800:STAT6 Transcription Factor / Q000378:metabolism*; D018418:Th2 Cells / Q000276:immunology / Q000378:metabolism
chemical_list: D039741:Antigens, Dermatophagoides; D016207:Cytokines; D058847:Inflammasomes; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D037742:Nanotubes, Carbon; D050800:STAT6 Transcription Factor; D007073:Immunoglobulin E; D020170:Caspase 1