title: Protease-dependent activation of epithelial cells by fungal allergens leads to morphologic changes and cytokine production.
abstract: BACKGROUND
  Proteases in extracts of Aspergillus fumigatus cause epithelial cell desquamation and release of proinflammatory cytokines.
  
  
  OBJECTIVE
  We sought to assess protease activity in Alternaria alternata, Cladosporium herbarum, and Aspergillus fumigatus extracts and study the ability of these extracts to cause desquamation and release of proinflammatory cytokines from epithelial cells.
  
  
  METHODS
  Protease activities of the fungal extracts were quantified. Changes with respect to cell morphology, cell desquamation, and cytokine production (IL-6 and IL-8) were measured in the absence and presence of the fungal extracts in an airway-derived epithelial cell line (A549) and primary epithelial nasal cells.
  
  
  RESULTS
  Fungal proteases differentially induced morphologic changes, cell desquamation, and production of IL-6 and IL-8 in a dose- and time-dependent fashion. Alternaria alternata extracts induced cell shrinking and cell desquamation and strongly enhanced the production of IL-6 and IL-8 at higher concentrations. Aspergillus fumigatus extracts caused cell shrinking, cell desquamation, and production of IL-6 and IL-8, even at low concentrations. The Aspergillus fumigatus-derived extract grown on collagen medium induced a strong dose-dependent decline in cytokine production at higher concentrations. Cladosporium herbarum extracts did not induce morphologic changes or cell desquamation but enhanced IL-6 and IL-8 productions at higher concentrations. The dependence of these effects on intact protease activity was shown by their abrogation by protease inhibitors.
  
  
  CONCLUSIONS
  Proteases present in fungal extracts interact with epithelial cells, leading to morphologic changes, cell desquamation, and induction of proinflammatory cytokines. It is proposed that these fungal proteases may activate epithelial cells through a protease-activated receptor type 2-driven mechanism.
authors.affiliation: Laboratory of Allergology and Pulmonology, Clinic for Internal Medicine, University Hospital, Groningen, The Netherlands.
authors.forename: H F
authors.initials: HF
authors.lastname: Kauffman
authors.forename: J F
authors.initials: JF
authors.lastname: Tomee
authors.forename: M A
authors.initials: MA
authors.lastname: van de Riet
authors.forename: A J
authors.initials: AJ
authors.lastname: Timmerman
authors.forename: P
authors.initials: P
authors.lastname: Borger
mesh_terms: D000485:Allergens / Q000494:pharmacology*; D000528:Alternaria / Q000276:immunology; D000946:Antigens, Fungal / Q000276:immunology* / Q000494:pharmacology; D001232:Aspergillus fumigatus / Q000276:immunology; D015550:Cell Degranulation / Q000187:drug effects; D002460:Cell Line; D002963:Cladosporium / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis*; D010450:Endopeptidases / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D009297:Nasal Mucosa / Q000166:cytology; D010936:Plant Extracts / Q000494:pharmacology
chemical_list: D000485:Allergens; D000946:Antigens, Fungal; D016207:Cytokines; D010936:Plant Extracts; D010450:Endopeptidases