title: Cross-matches on donor cadaver retinal pigment epithelial cells in corneal risk patients.
abstract: BACKGROUND
  Allografts can be rejected either through the antibody-mediated or cellular pathways. The objective of this study was to look at the extent of antibody formation in patients awaiting re-keratoplasty using cross-matches on cadaver retinal pigment epithelial (RPE) cells.
  
  
  METHODS
  Cadaver RPE cells were derived by trypsin digestion from donor eyes (n = 1200). After 3 days of cell cultivation, the cells were adherent and began to lose their pigment. By day 7 most cells were clear and grew as a polygonal monolayer. MHC class I expression by RPE cells was studied by the W6/32 (anti-HLA-A, B, C) monoclonal antibody (MoAb) and that of class II (HLA-DR) by the 136 MoAb. Normal RPE cells express few class I and no detectable class II antigens. For the induction of MHC expression, cells were subsequently stimulated with 250 U/ml of recombinant gamma-interferon for 5 days. Cells were used for tissue typing and also for cross-matches with recipient serum. Cross-matches were subsequently performed and measured by flow cytometry.
  
  
  RESULTS
  Both class I and class II antigens were strongly enhanced, as could be shown by immunohistochemical staining. Some 20% of those patients awaiting rekeratoplasty (n = 60) were positive for anti-HLA antibodies. In one case anti-DR3 antibodies were detected in a recipient who had had several rejection episodes after keratoplasty.
  
  
  CONCLUSIONS
  RPE cells are not only useful for cadaver post-mortem HLA typing but also for donor-specific cross-matches. The degree of antibody formation after keratoplasty in rejecting patients was, however, low. This may imply that anti-HLA antibodies are not the major cause of corneal graft loss after keratoplasty.
authors.affiliation: Institute of Immunology, University of Kiel, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Zavazava
authors.forename: B
authors.initials: B
authors.lastname: Nölle
authors.forename: G
authors.initials: G
authors.lastname: Duncker
authors.forename: S
authors.initials: S
authors.lastname: Jenisch
authors.forename: E
authors.initials: E
authors.lastname: Westphal
authors.forename: V
authors.initials: V
authors.lastname: Eckstein
authors.forename: W
authors.initials: W
authors.lastname: Müller-Ruchholtz
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D000917:Antibody Formation / Q000276:immunology; D002102:Cadaver; D002478:Cells, Cultured; D003315:Cornea / Q000276:immunology*; D005434:Flow Cytometry; D006084:Graft Rejection / Q000276:immunology*; D006085:Graft Survival / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000032:analysis* / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000032:analysis* / Q000276:immunology; D006650:Histocompatibility Testing / Q000379:methods; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D015948:Keratoplasty, Penetrating / Q000276:immunology*; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D011994:Recombinant Proteins; D012086:Reoperation; D012307:Risk Factors; D014019:Tissue Donors; D014184:Transplantation, Homologous
chemical_list: D000911:Antibodies, Monoclonal; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D011994:Recombinant Proteins; D007371:Interferon-gamma