title: Gamma-irradiation reduces the allogenicity of donor corneas.
abstract: OBJECTIVE
  To evaluate the utility and allogenicity of gamma-irradiated corneal allografts.
  
  
  METHODS
  Corneal buttons were harvested from C57BL/6 mice and decellularized with gamma irradiation. Cell viability was assessed using TUNEL and viability/cytotoxicity assays. Orthotopic penetrating keratoplasty was performed using irradiated or nonirradiated (freshly excised) C57BL/6 donor grafts and BALB/c or C57BL/6 recipients. Graft opacity was assessed over an 8-week period and graft survival was evaluated using Kaplan-Meier survival curves. Mixed-lymphocyte reactions and delayed-type hypersensitivity assays were performed to evaluate T-cell alloreactivity. Real-time PCR was used to investigate the corneal expression of potentially pathogenic T-helper 1, 2, and 17 cell-associated cytokines.
  
  
  RESULTS
  Corneal cells were devitalized by gamma irradiation as evidenced by widespread cellular apoptosis and plasma membrane disruption. Nonirradiated allograft and isograft rates of survival were superior to irradiated allograft and isograft rates of survival (P < 0.001). Mixed lymphocyte reactions demonstrated that T-cells from irradiated allograft recipients did not exhibit a secondary alloimmune response (P < 0.001). Delayed-type hypersensitivity assays demonstrated that irradiated allografts did not elicit an alloreactive delayed-type hypersensitivity response in graft recipients (P ≤ 0.01). The corneal expression of T-helper 1, 2, and 17 cell-associated cytokines was significantly lower in failed irradiated allografts than rejected nonirradiated allografts (P ≤ 0.001).
  
  
  CONCLUSIONS
  Gamma-irradiated corneas failed to remain optically clear following murine penetrating keratoplasty; however, gamma irradiation reduced the allogenicity of these corneas, potentially supporting their use in procedures such as anterior lamellar keratoplasty or keratoprosthesis implantation.
authors.affiliation: Schepens Eye Research Institute, Massachusetts Eye & Ear Infirmary, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: William
authors.initials: W
authors.lastname: Stevenson
authors.forename: Sheng-Fu
authors.initials: SF
authors.lastname: Cheng
authors.forename: Parisa
authors.initials: P
authors.lastname: Emami-Naeini
authors.forename: Jing
authors.initials: J
authors.lastname: Hua
authors.forename: Eleftherios I
authors.initials: EI
authors.lastname: Paschalis
authors.forename: Reza
authors.initials: R
authors.lastname: Dana
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Saban
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000276:immunology / Q000528:radiation effects; D003315:Cornea / Q000276:immunology* / Q000528:radiation effects*; D003318:Corneal Opacity / Q000209:etiology* / Q000276:immunology; D005720:Gamma Rays / Q000009:adverse effects / Q000627:therapeutic use*; D006084:Graft Rejection / Q000276:immunology / Q000517:prevention & control; D006085:Graft Survival / Q000276:immunology / Q000528:radiation effects; D007165:Immunosuppression Therapy / Q000379:methods; D007519:Isoantigens / Q000276:immunology; D015948:Keratoplasty, Penetrating / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology / Q000528:radiation effects; D014181:Transplantation Immunology / Q000528:radiation effects*; D014184:Transplantation, Homologous
chemical_list: D007519:Isoantigens