Abstract
Abstract: :
Purpose: To characterize large–scale gene expression patterns in peripheral blood from patients undergoing corneal transplantation and extend our understanding of the molecular changes underlying corneal graft rejection, using oligonucleotide microarrays. Methods:Atlas Plastic Human 12K Microarrays containing 12,000 gene–specific oligonucleotides were hybridised with first strand 33P–labelled cDNA generated from total RNA in the peripheral blood from patients who underwent corneal transplantation with graft rejection (n=4) and patients whose grafts were not rejected (n=4). The data was analysed for differential gene expression, which will be confirmed by real–time RT–PCR and Northern blotting for selected genes. Results:A total of one hundred and forty–eight genes showed at least a two–fold change in expression. Among them, 100 genes were up–regulated, whereas 48 genes were down–regulated in the blood samples from rejecting patients compared with non–rejecting patients. Approximately 25% of these genes are currently functionally unclassified. The identified differentially expressed genes with a known function included cytokines and inflammatory mediators, transcription related proteins, extracellular transport proteins, intracellular transducers, and cell surface antigens. Conclusions: The cDNA Microarray technology has been demonstrated to be a useful tool to monitor the expression of a large number of genes simultaneously. Most results have to be considered to be preliminary as there is a major challenge to develop an accepted approach to validate the obtained mass of data. However, our data imply that multiple pathways might be involved in the graft rejection of corneal transplantation, including immune and non–immune response.
Keywords: cornea: clinical science • transplantation