Feasibility of Diagnosing Subclinical Renal Allograft Rejection in Children By Whole Blood Gene Expression Analysis
Feasibility of Diagnosing Subclinical Renal Allograft Rejection in Children By Whole Blood Gene Expression Analysis
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DOI：
10.1097/tp.0b013e3181883fb0
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发表时间：
2008-11-15
期刊：
影响因子：
6.2
通讯作者：
Laine, Jarmo
中科院分区：
文献类型：
--
作者：
Alakulppi, Noora;Seikku, Paula;Laine, Jarmo
背景方案活检用于监测移植后的同种异体移植物组织学。然而，活检是一种具有潜在并发症的侵入性手术，需要特殊的设施，并且对于移植物的重复监测是不切实际的。一种非侵入性、稳健且快速的诊断方法将受到欢迎。从血液样品中监测基因表达可以提供这样的手段。对31例儿童肾移植受者在3个月或6个月方案活检时采集的全血样本进行了研究，其中13例经活检证实为亚临床排斥反应（SCR）。将样品收集到含有RNA稳定化试剂的试管中，使得能够在正常病房时间表期间进行可行的收集。在所有患者中，检测候选基因CD 154和诱导性T细胞共刺激因子（ICOS）的基因表达。采用实时定量PCR（RT-QPCR）技术，对10例患者的90个免疫相关基因进行了低密度芯片检测。此外，还对8例患者进行了全基因组微阵列分析。CD 154和ICOS基因表达均不能诊断SCR（CD 154和ICOS的中位表达水平分别为1.25 vs. 1.16和1.95 vs. 1.61）。此外，低密度阵列上没有一个基因的表达水平与SCR相关。最后，在芯片分析中，在SCR和正常患者的基因表达之间没有发现差异可以用RT-QPCR在17个基因中验证。在我们相对较小的系列研究中，没有发现可靠的SCR全血基因表达生物标志物，需要进一步研究以确定表达的微小变化是否可以提供支持性诊断方法。
Background. Protocol biopsies are used to monitor allograft histology after transplantation. However, biopsy is an invasive procedure with potential complications, requires special facilities, and is unpractical for repeated monitoring of the graft. A noninvasive, robust, and rapid diagnostic method would be welcomed. Monitoring gene expression from blood samples could provide such a means.Methods. Whole blood samples taken at the time of 3- or 6-month protocol biopsy in 31 pediatric renal transplant recipients, 13 of whom had biopsy-proven subclinical rejection (SCR), were studied. The samples were collected into tubes containing an RNA stabilization reagent enabling feasible collection during a normal ward schedule. In all patients, the gene expression of candidate genes CD154 and inducible T-cell co-stimulator (ICOS) was measured. A low-density array containing 90 immunologic-related genes were measured with real-time quantitative PCR (RT-QPCR) in 10 patients. In addition, a whole genome microarray analysis was performed in eight patients.Results. Neither CD154 nor ICOS gene expression was diagnostic for SCR (median expression level 1.25 vs. 1.16 and 1.95 vs. 1.61 for CD154 and ICOS, respectively). In addition, expression levels of none of the genes on the low-density array were associated with SCR. Finally, in the microarray analysis none of the found differences between SCR and normal patients' gene expression could be validated with RT-QPCR in 17 genes.Conclusions. In our relatively small series no robust whole blood gene expression biomarker for SCR was found. Further studies are needed to determine whether small changes in expression may provide a supporting diagnostic method.