title: Cytokine deposition alters leukocyte morphology and initial recruitment of monocytes and γδT cells after corneal injury.
abstract: OBJECTIVE
  An in vivo mouse model reproducibly induces recurrent epithelial erosions in wild-type mice spontaneously 2 weeks after a single 1.5-mm corneal debridement wound made with a dulled blade. When 1.5-mm wounds are made by a rotating burr so that the corneal epithelial basement membrane is removed, corneas heal without developing erosions. Here, we characterize differences in cytokine deposition and changes in leukocytes between 0 and 6 hours after dulled-blade and rotating-burr wounding.
  
  
  METHODS
  BALB/c mice were used to study 1.5-mm corneal wounds made using a dulled blade or a rotating burr. Mice were studied immediately after wounding (0 hour) and at 6 hours in vivo and in vitro in organ culture. Corneas, corneal extracts, and collagenase digests from naïve and wounded mice were used for three-dimensional (3D) confocal imaging, cytokine arrays, and flow cytometry.
  
  
  RESULTS
  Confocal imaging showed CD45, a protein derived from leukocytes, accumulates at the wound edge by 3 and 6 hours after wounding in vivo but not in vitro with more CD45 accumulating after dulled-blade compared with rotating-burr wounds. Morphologic changes occurred in CD45+ leukocytes and higher levels for several cytokines were detected in the stromal wound bed within minutes following dulled-blade wounds. Flow cytometry showed significantly more monocytes (CD45+/CD11b+/Ly6C+) and γδT cells (CD45+/GL3+) recruited into the corneas of mice with dulled-blade wounds by 6 hours.
  
  
  CONCLUSIONS
  Differences in cytokine-driven leukocyte responses are seen after dulled-blade debridement compared with rotating-burr injury.
authors.affiliation: Department of Anatomy and Regenerative Biology and Department of Ophthalmology, The George Washington University Medical School, Washington, DC, United States.
authors.forename: Sonali
authors.initials: S
authors.lastname: Pal-Ghosh
authors.forename: Ahdeah
authors.initials: A
authors.lastname: Pajoohesh-Ganji
authors.forename: A Sue
authors.initials: AS
authors.lastname: Menko
authors.forename: Hye-young
authors.initials: HY
authors.lastname: Oh
authors.forename: Gauri
authors.initials: G
authors.lastname: Tadvalkar
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Saban
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Stepp
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000378:metabolism / Q000473:pathology; D003315:Cornea / Q000276:immunology / Q000473:pathology; D065306:Corneal Injuries; D016207:Cytokines / Q000378:metabolism*; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000293:injuries / Q000378:metabolism / Q000473:pathology; D005131:Eye Injuries / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D007113:Immunity, Innate; D007962:Leukocytes / Q000473:pathology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D009000:Monocytes / Q000473:pathology; D014945:Wound Healing / Q000276:immunology*
chemical_list: D016207:Cytokines
keywords: basement membrane; innate immune response; monocytes; mouse; γδT cells