PDF(Pubmed)
Abstract：
Zero-time biopsies are taken to determine the quality of the donor organ at the time of transplantation. Histological analyses alone have so far not been able to identify parameters that allow the prediction of subsequent rejection episodes or graft survival. This study investigated whether gene expression analyses of zero-time biopsies might support this prediction. Using a well-characterized cohort of 26 zero-time biopsies from renal transplant patients that include 4 living donor (LD) and 22 deceased donor (DD) biopsies that later developed no rejection (Ctrl, n = 7), delayed graft function (DGF, n = 4), cellular (T-cell mediated rejection; TCMR, n = 8), or antibody-mediated rejection (ABMR, n = 7), we analyzed gene expression profiles for different types of subsequent renal transplant complication. To this end, RNA was isolated from formalin-fixed, paraffin-embedded (FFPE) sections and gene expression profiles were quantified. Results were correlated with transplant data and B-cell, and plasma cell infiltration was assessed by immunofluorescence microscopy. Both principal component analysis and clustering analysis of gene expression data revealed marked separation between LDs and DDs. Differential expression analysis identified 185 significant differentially expressed genes (adjusted p < 0.05). The expression of 68% of these genes significantly correlated with cold ischemia time (CIT). Furthermore, immunoglobulins were differentially expressed in zero-time biopsies from transplants later developing rejection (TCMR + ABMR) compared to non-rejected (Ctrl + DGF) transplants. In addition, immunoglobulin expression did not correlate with CIT but was increased in transplants with previous acute renal failure (ARF). In conclusion, gene expression profiles in zero-time biopsies derived from LDs are markedly different from those of DDs. Pre-transplant ARF increased immunoglobulin expression, which might be involved in triggering later rejection events. However, these findings must be confirmed in larger cohorts and the role of early immunoglobulin upregulation in zero-biopsies needs further clarification.
摘要：
进行零时间活检以确定移植时供体器官的质量。迄今为止，仅组织学分析还不能确定允许预测后续排斥事件或移植物存活的参数。这项研究调查了零时活检的基因表达分析是否可能支持这一预测。使用来自肾移植患者的26个零时活检的特征队列，其中包括4个活体供体（LD）和22个死者供体（DD）活检，这些活检后来没有发生排斥反应（Ctrl，n=7),延迟的移植物功能(DGF，n=4),细胞(T细胞介导的排斥反应；TCMR，n=8),或抗体介导的排斥反应(ABMR，n=7),我们分析了不同类型的后续肾移植并发症的基因表达谱.为此,从福尔马林固定的RNA中分离，对石蜡包埋(FFPE)切片和基因表达谱进行定量。结果与移植数据和B细胞相关，和浆细胞浸润通过免疫荧光显微镜评估。基因表达数据的主成分分析和聚类分析均显示LD和DD之间存在明显的分离。差异表达分析鉴定了185个显著差异表达的基因(调整的p<0.05)。这些基因的68％的表达与冷缺血时间（CIT）显着相关。此外，与非排斥(Ctrl+DGF)移植相比,免疫球蛋白在随后发生排斥反应的移植(TCMR+ABMR)的零时活检中差异表达.此外,免疫球蛋白表达与CIT不相关,但在既往急性肾衰竭(ARF)的移植中增加.总之,来自LD的零时间活检中的基因表达谱与DD的基因表达谱明显不同。移植前ARF增加免疫球蛋白表达，这可能涉及触发以后的拒绝事件。然而,这些发现必须在更大的队列中得到证实,早期免疫球蛋白上调在零活检中的作用需要进一步澄清.
