title: Corneal graft rejection occurs despite Fas ligand expression and apoptosis of infiltrating cells.
abstract: OBJECTIVE
  Constitutive expression of Fas ligand (CD95L) protects the eye against cell mediated immune responses by inducing apoptosis in infiltrating Fas bearing T cells. This study was designed to examine Fas ligand expression on acutely rejecting rat corneal grafts and to investigate the kinetics of induction of apoptosis in infiltrating leucocytes.
  
  
  METHODS
  Orthotopic penetrating corneal transplantation was performed between genetically disparate inbred rats. Fas ligand expression and the phenotype of infiltrating leucocytes were examined by immunohistochemistry. Apoptotic nuclei were visualised in sections of normal rat cornea, rejecting allografts, and time matched isografts by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labelling (TUNEL) and quantified by video image analysis. Staining with Hoechst dye 33258 was used to confirm the presence of apoptotic nuclei.
  
  
  RESULTS
  Fas ligand was expressed on corneal endothelial and epithelial cells during acute corneal graft rejection. At all time points examined, including as early as the fifth postoperative day, the cells infiltrating both corneal isografts and allografts were TUNEL positive. By the 15th postoperative day, over 90% of all nuclei, many of which were T cells, were apoptotic.
  
  
  CONCLUSIONS
  Expression of Fas ligand is not downregulated on the cornea during allograft rejection and infiltrating leucocytes in both isografts and allografts die rapidly in situ. Despite the death of the cells believed to be responsible for rejection, isografts survive indefinitely whereas allografts are irreparably damaged.
authors.affiliation: Department of Ophthalmology, Flinders Medical Centre, Bedford Park, SA 5042, Australia. keryn.williams@flinders.edu.au
authors.forename: K A
authors.initials: KA
authors.lastname: Williams
authors.forename: S D
authors.initials: SD
authors.lastname: Standfield
authors.forename: J R
authors.initials: JR
authors.lastname: Smith
authors.forename: D J
authors.initials: DJ
authors.lastname: Coster
mesh_terms: D000208:Acute Disease; D000818:Animals; D017209:Apoptosis; D016039:Corneal Transplantation / Q000473:pathology*; D004728:Endothelium, Corneal / Q000378:metabolism / Q000473:pathology; D019573:Epithelium, Corneal / Q000378:metabolism / Q000473:pathology; D053222:Fas Ligand Protein; D005260:Female; D006084:Graft Rejection / Q000378:metabolism / Q000473:pathology*; D007124:Immunoenzyme Techniques; D020287:In Situ Nick-End Labeling; D008024:Ligands; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D011920:Rats, Inbred WF
chemical_list: D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008024:Ligands; D008562:Membrane Glycoproteins