Department of Medical Genetics, University of Manchester, UK.
Eye (Lond). 2009 Feb;23(2):461-9. doi: 10.1038/eye.2008.306. Epub 2008 Oct 10.
Allograft rejection is the main cause of graft failure in human corneal transplantation, for which underlying pathomechanism is not yet clear. We compared gene expression in the peripheral blood of patients who after undergoing corneal transplantation experienced graft rejection with those patients who accepted grafts.
Sixty-six patients who underwent corneal transplantation were studied including 18 patients who suffered subsequent graft rejection. cDNA array technology was used to survey and quantify transcript expression. A semiquantitative reverse transcriptase-PCR (RT-PCR) was used to confirm the gene expression pattern measured by a cDNA array of selected genes.
Among 265 genes present on the array, eight genes were found to be differentially expressed. Four genes (Rac 2, RhoA, paxillin, and CD18) were further analysed by semiquantitative RT-PCR, and significant differences in mRNA expression levels in the rejection group were confirmed.
Our study demonstrated that the expression of Rac2 mRNA was upregulated in the peripheral blood of patients experiencing corneal transplantation rejection compared to those patients who had no rejection episodes. In addition, three genes, RhoA, paxillin, and CD18, showed decreased expression in rejecting patients. cDNA array technology provides a potentially useful approach to identify novel genes that might participate in pathogenic pathways during corneal graft rejection.
同种异体移植物排斥是导致人类角膜移植失败的主要原因,但其潜在的发病机制尚不清楚。我们比较了接受角膜移植后发生排斥反应的患者和接受移植的患者外周血中的基因表达。
研究了 66 例接受角膜移植的患者,其中 18 例随后发生移植物排斥反应。使用 cDNA 微阵列技术检测和定量转录物表达。半定量逆转录聚合酶链反应(RT-PCR)用于确认通过 cDNA 微阵列测量的选定基因的基因表达模式。
在存在于阵列上的 265 个基因中,发现有 8 个基因表达存在差异。进一步通过半定量 RT-PCR 分析了 4 个基因(Rac2、RhoA、桩蛋白和 CD18),证实了排斥组中 mRNA 表达水平的显著差异。
我们的研究表明,与没有排斥反应的患者相比,经历角膜移植排斥反应的患者外周血中 Rac2 mRNA 的表达上调。此外,排斥患者的 3 个基因 RhoA、桩蛋白和 CD18 表达下调。cDNA 微阵列技术为鉴定可能参与角膜移植物排斥过程中致病途径的新基因提供了一种潜在有用的方法。