Cytokine deposition alters leukocyte morphology and initial recruitment of monocytes and γδT cells after corneal injury

Invest Ophthalmol Vis Sci. 2014 Apr 28;55(4):2757-65. doi: 10.1167/iovs.13-13557.


Purpose: 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.

Keywords: basement membrane; innate immune response; monocytes; mouse; γδT cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Basement Membrane / metabolism
  • Basement Membrane / pathology
  • Cornea / immunology
  • Cornea / pathology
  • Corneal Injuries*
  • Cytokines / metabolism*
  • Disease Models, Animal
  • Epithelium, Corneal / injuries
  • Epithelium, Corneal / metabolism
  • Epithelium, Corneal / pathology
  • Eye Injuries / immunology*
  • Eye Injuries / metabolism
  • Eye Injuries / pathology
  • Immunity, Innate*
  • Leukocytes / pathology*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Confocal
  • Monocytes / pathology
  • Wound Healing / immunology*


  • Cytokines