title: Identification of Interleukin-1β-Producing Monocytes That Are Susceptible to Pyronecrotic Cell Death in Patients With Neonatal-Onset Multisystem Inflammatory Disease.
abstract: OBJECTIVE
  Spontaneous inflammatory responses initiated by NLRP3 mutations promote inflammasome-mediated interleukin-1β (IL-1β) processing and release and can induce rapid necrotic cell death. The cells that produce IL-1β in neonatal-onset multisystem inflammatory disease (NOMID) have not been clearly identified, nor have the mechanisms mediating IL-1β release and cell death been completely elucidated.
  
  
  METHODS
  Whole blood cells were stimulated with lipopolysaccharide (LPS) in the presence of cathepsin B and caspase 1 inhibitors, followed by ATP treatment. Supernatants were collected and incubated with IL-1β-capturing beads. Cells were fixed, permeabilized, and stained for a combination of cell surface and intracellular markers, and a novel flow cytometry bead-based assay was used to measure secreted IL-1β. LPS-stimulated cells were also evaluated using immunofluorescence microscopy.
  
  
  RESULTS
  Monocytes characterized by CD14(high) -CD16(low) expression and intracellular CD83 were increased in NOMID patients and were responsible for the majority of IL-1β production in response to LPS stimulation. This population of monocytes also underwent a rapid death response with LPS alone that is temporally associated with IL-1β and ASC release and has characteristic features of pyronecrotic but not pyroptotic cell death. Inhibition of cell death reduced IL-1β production from NOMID patient cells. In addition, IL-1 triggered cell death in monocytes from NOMID patients.
  
  
  CONCLUSIONS
  Our findings indicate that monocytes are the predominant IL-1β-producing cell population in the peripheral blood of NOMID patients. Furthermore, they suggest that IL-1 receptor blockade may work in part by preventing pyronecrotic cell death, which may be an important target in NOMID and other forms of cryopyrin-associated periodic syndromes.
authors.affiliation: National Institute of Arthritis and Musculoskeletal and Skin Diseases, NIH, Bethesda, Maryland.
authors.forename: Jehad H
authors.initials: JH
authors.lastname: Edwan
authors.affiliation: National Institute of Arthritis and Musculoskeletal and Skin Diseases, NIH, Bethesda, Maryland.
authors.forename: Raphaela
authors.initials: R
authors.lastname: Goldbach-Mansky
authors.affiliation: National Institute of Arthritis and Musculoskeletal and Skin Diseases, NIH, Bethesda, Maryland.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Colbert
mesh_terms: D000293:Adolescent; D000328:Adult; D015703:Antigens, CD / Q000276:immunology; D002352:Carrier Proteins / Q000235:genetics; D061945:Caspase Inhibitors / Q000494:pharmacology; D002401:Cathepsin B / Q000037:antagonists & inhibitors; D016923:Cell Death / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D056587:Cryopyrin-Associated Periodic Syndromes / Q000235:genetics / Q000276:immunology*; D005260:Female; D058851:GPI-Linked Proteins / Q000276:immunology; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D007223:Infant; D053583:Interleukin-1beta / Q000276:immunology*; D018950:Lipopolysaccharide Receptors / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology; D009000:Monocytes / Q000187:drug effects / Q000276:immunology*; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D009336:Necrosis / Q000276:immunology*; D000069292:Pyroptosis; D017452:Receptors, IgG / Q000276:immunology; D055815:Young Adult; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D002352:Carrier Proteins; D061945:Caspase Inhibitors; D002401:Cathepsin B; D058851:GPI-Linked Proteins; D007136:Immunoglobulins; D053583:Interleukin-1beta; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D017452:Receptors, IgG; D000097802:CD83 Antigen; C428200:FCGR3B protein, human; C508585:IL1B protein, human; C453881:NLRP3 protein, human