Abstract
Background: :
Lactoferrin deposites at secondary corneal amyloidosis. It has been described that lactoferrin in eyes is mainly derived from the lacrimal gland. However, lactoferrin is found to deposit at deeper layers of the corneal epithelium in secondary amyloidosis. This observation led us to investigate other possible sources of lactoferrin other than the lacrimal gland.
Purpose: :
To investigate the production source of lactoferrin deposited at secondary corneal amyloidosis in vitro and in vivo.
Methods: :
In vitro experiments, human corneal epithelial cells (cell line HCE–T) were applied to determine lactoferrin expression in terms of immuno–histochemistry and Reverse Transcription–Polymerase Chain Reaction (RT–PCR). The cultured cells were fixed at confluence and 48 hours after the wounding by a scraper, and were subject to immuno–staining. For RT–PCR test, mRNA was extracted at 0,24,48,72,120 (at wound closure) hours after the wounding, and 24 hours after wound closure. In vivo experiments, lactoferrin deposition was analyzed by immuno–staining in the corneal specimens obtained from keratoplasty (four keratoconus without amyloidsis and four controls).
Results: :
The cultured human corneal epithelial cells (cell line HCE–T) expressed detectable level of lactoferrin mRNA and the protein product during the wound healing process. In human specimens, lactoferrin–positive staining was observed at the basal layer of the corneal epithelium in two out of four keratoconus without amyloidosis.
Conclusions: :
Corneal epithelial cell serves as a putative source of lactoferrin deposited at the secondary corneal amyloidosis.
Keywords: cornea: basic science • pathology: experimental