title: A Pseudomonas aeruginosa strain isolated from a contact lens-induced acute red eye (CLARE) is protease-deficient.
abstract: OBJECTIVE
  Pseudomonas aeruginosa proteases are thought to be important virulence factors in the pathogenesis of corneal disease. This study examined protease production from two strains of P. aeruginosa responsible for two very distinct clinical diseases: strain Paer1, isolated from a Contact Lens-induced Acute Red Eye (CLARE), and strain KEI 1025, isolated from a corneal ulcer. Strains were compared to a laboratory strain (ATCC 19660) known to produce severe keratitis in experimentally infected mice for protease production and for ocular virulence.
  
  
  METHODS
  Protease production was examined with colorimetric assays, gelatin zymography and western blots. Elastase A activity was quantitated with a staphylolytic assay. Ocular virulence was examined using a mouse scratch model of keratitis.
  
  
  RESULTS
  In contrast to strains KEI 1025 or ATCC 19660, Paer1 was unable to produce enzymatically active elastase A, elastase, and protease IV. All three strains produced active alkaline protease. Strains KEI 1025 and ATCC 19660 produced a fulminant keratitis in mice whereas Paer1 produced a mild transient infection. Restoration of elastase activity in Paer1 via genetic complementation did not result in a virulent phenotype. Co-infection of mouse eyes with strains Paer1 and ATCC 19660 resulted in the eventual loss of Paer1 from corneal tissue.
  
  
  CONCLUSIONS
  These studies suggest that P. aeruginosa elastase A and/or protease IV, but not alkaline protease or elastase, contribute to the ocular virulence of this organism.
authors.affiliation: Department of Immunology/Microbiology, Wayne State University, School of Medicine, Detroit, MI 48201, USA.
authors.forename: P S
authors.initials: PS
authors.lastname: Estrellas
authors.forename: L G
authors.initials: LG
authors.lastname: Alionte
authors.forename: J A
authors.initials: JA
authors.lastname: Hobden
mesh_terms: D000208:Acute Disease; D000818:Animals; D001769:Blood; D002455:Cell Division / Q000187:drug effects; D015169:Colony Count, Microbial; D003261:Contact Lenses / Q000009:adverse effects*; D003320:Corneal Ulcer / Q000209:etiology / Q000382:microbiology*; D003470:Culture Media / Q000494:pharmacology; D020689:Exopeptidases / Q000378:metabolism*; D005123:Eye / Q000382:microbiology; D005260:Female; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008826:Microbial Sensitivity Tests; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000302:isolation & purification / Q000472:pathogenicity; D014774:Virulence
chemical_list: D003470:Culture Media; D020689:Exopeptidases