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Published on: April 11, 2011
Expression of MHC class I and II molecules by cadaver retinal pigment epithelium cells: optimization of post-mortem
N Zavazava1, M Halene, E Westphal
1Department of Immunology, University of Kiel, Germany.
Insights
Retinal pigment epithelium (RPE) cells express MHC antigens after interferon-gamma (IFN-gamma) stimulation. This finding improves post-mortem HLA typing for better organ graft matching and survival rates.
Area of Science:
- Immunology
- Ophthalmology
- Transplantation Immunology
Background:
- Retinal pigment epithelium (RPE) cells play a crucial role in ocular immunity.
- Accurate human leukocyte antigen (HLA) typing is vital for successful organ transplantation.
- Current cadaver HLA typing methods have limitations, especially for organs preserved long after death.
Purpose of the Study:
- To investigate the expression of major histocompatibility complex (MHC) antigens on RPE cells.
- To assess the effect of interferon-gamma (IFN-gamma) on RPE cell MHC antigen expression.
- To improve post-mortem HLA typing techniques for enhanced graft survival.
Main Methods:
- RPE cells were stimulated with varying concentrations of IFN-gamma.
- MHC antigen expression (Class I, HLA-DR, HLA-DP, HLA-DQ) was quantified.
- Serological typing using standard lymphocytotoxicity assay and 1D-IEF was performed on stimulated RPE cells.
Main Results:
- IFN-gamma induced significant expression of MHC Class I and HLA-DR on RPE cells.
- Specific concentrations of IFN-gamma modulated the expression of HLA-DP and HLA-DQ.
- High concordance was observed between RPE cell typing and standard lymphocytotoxicity tests.
Conclusions:
- RPE cells can be effectively typed for MHC antigens post-mortem.
- This method enhances the accuracy of HLA typing for organs like the cornea.
- Improved post-mortem typing facilitates better graft matching and increases graft survival rates.
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.
