Abstract
The objective of this study was to investigate the expression of MHC antigens by retinal pigment epithelium cells (RPE) after stimulation with interferon-gamma (IFN-gamma) and to improve the currently practised technique of cadaver HLA typing. A concentration of 100 U/ml IFN-gamma induced expression of class I molecules up to greater than 90% 3 days after stimulation, whereas 50 U/ml were required for the expression of HLA-DR to greater than 90%. A concentration of 750 U/ml induced 35-45% expression of HLA-DP and less than 25% HLA-DQ after 3 days. Cells were serologically typed using the standard lymphocytotoxicity assay 3 days after stimulation with 250 U/ml IFN-gamma. Typing of class I specificities was complemented by one-dimensional isoelectric focusing (1D-IEF). We observed high concordance between the results of the RPE typing and the lymphocytotoxicity test on the same donors. Our results show complete typing of class I and II antigens post-mortem, which, in particular, enables graft matching and improvement of graft survival in recipients of organs removed many hours after death such as the cornea.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baumgartner I., Huber-Spitzy V., Grabner G., Mayr W. R. HLA typing from human donor eyes. Graefes Arch Clin Exp Ophthalmol. 1989;227(6):541–543. doi: 10.1007/BF02169449. [DOI] [PubMed] [Google Scholar]
- Baumgartner I., Mayr W. R., Grabner G. Corneal grafting--strategy for optimal results. Tissue Antigens. 1989 Jul;34(1):5–8. doi: 10.1111/j.1399-0039.1989.tb01710.x. [DOI] [PubMed] [Google Scholar]
- Dobbe L. M., Stam N. J., Neefjes J. J., Giphart M. J. Biochemical complexity of serum HLA class I molecules. Immunogenetics. 1988;27(3):203–210. doi: 10.1007/BF00346587. [DOI] [PubMed] [Google Scholar]
- FAURE J. P. LES R'EACTIONS IMMUNOLOGIQUES DANS LES GREFFES DE LA CORN'EE. Arch Ophtalmol Rev Gen Ophtalmol. 1964 Sep;24:501–CONTD. [PubMed] [Google Scholar]
- Flood M. T., Gouras P., Kjeldbye H. Growth characteristics and ultrastructure of human retinal pigment epithelium in vitro. Invest Ophthalmol Vis Sci. 1980 Nov;19(11):1309–1320. [PubMed] [Google Scholar]
- Liversidge J. M., Sewell H. F., Forrester J. V. Human retinal pigment epithelial cells differentially express MHC class II (HLA, DP, DR and DQ) antigens in response to in vitro stimulation with lymphokine or purified IFN-gamma. Clin Exp Immunol. 1988 Sep;73(3):489–494. [PMC free article] [PubMed] [Google Scholar]
- MAUMENEE A. E. The influence of donor-recipient sensitization on corneal grafts. Am J Ophthalmol. 1951 May;34(5 2):142–152. doi: 10.1016/0002-9394(51)90019-0. [DOI] [PubMed] [Google Scholar]
- Neefjes J. J., Doxiadis I., Stam N. J., Beckers C. J., Ploegh H. L. An analysis of class I antigens of man and other species by one-dimensional IEF and immunoblotting. Immunogenetics. 1986;23(3):164–171. doi: 10.1007/BF00373817. [DOI] [PubMed] [Google Scholar]
- Stam N. J., Spits H., Ploegh H. L. Monoclonal antibodies raised against denatured HLA-B locus heavy chains permit biochemical characterization of certain HLA-C locus products. J Immunol. 1986 Oct 1;137(7):2299–2306. [PubMed] [Google Scholar]
- TERASAKI P. I., MCCLELLAND J. D. MICRODROPLET ASSAY OF HUMAN SERUM CYTOTOXINS. Nature. 1964 Dec 5;204:998–1000. doi: 10.1038/204998b0. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]