Grigoryev D N, Liu M, Cheadle C, Barnes K C, Rabb H
Division of Clinical Immunology, Johns Hopkins University, Baltimore, Maryland, USA.
Transplant Proc. 2006 Dec;38(10):3333-6. doi: 10.1016/j.transproceed.2006.10.129.
Increased organ ischemia time leads to delayed graft function (DGF), increased acute rejection (AR), enhanced chronic allograft nephropathy (CAN), and reduced long-term allograft survival. The mechanisms by which IRI predisposes to AR and CAN are unknown. We hypothesized that gene expression profiling of ischemia-reperfusion injury (IRI)-affected kidney would identify how IRI predisposes to AR and CAN. Furthermore, we examined how current immunosuppressive drug molecular targets are altered by IRI. C57BL/6J mice were exposed to 30 (n = 3) or 60 (n = 3) minutes of bilateral kidney ischemia or sham surgery (n = 5). At 36 hour kidney tissue was collected and analyzed using Affymetrix 430MOEA (22626 genes) array and GC-RMA-SAM pipeline. Genes with the false discovery rate (q < 1%) and +/-50% fold change (FC) were considered affected by IRI. Genes coding for histocompatibility and antigen-presenting factors, calcineurin, and mammalian target of rapamycin (mTOR) pathway-associated proteins were selected using Gene Ontology (GO) analysis. GO analysis identified 10 and 17 alloimmunity-related genes affected by IRI induced by 30 and 60 minutes of ischemia, respectively, including Traf6 (FC = 2.99) and H2-D1 (FC = 2.58). We also detected significant IRI genomic responses in calcineurin and mTOR pathways represented by Fkbp5 (FC = 4.18) and Fkbp1a (FC = 2.0), and Eif4ebp1 (FC = 16.8) and Akt1 (FC = 3.64), respectively. These data demonstrated that IRI up-regulates expression of several alloimmunity-associated genes, which can in turn enhance alloimune responses. Our discovery of IRI-induced up-regulation of genes associated with calcineurin and mTOR pathways are consistent with clinical observations that FK506 and Rapamycin can alter the course of DGF. Further validation and dissection of these pathways can lead to novel approaches by which improved management of early "nonimmune" transplant events can decrease susceptibility to more classic "immune" changes and CAN.
器官缺血时间延长会导致移植肾功能延迟恢复（DGF）、急性排斥反应（AR）增加、慢性移植肾肾病（CAN）加重以及移植肾长期存活时间缩短。缺血再灌注损伤（IRI）易引发AR和CAN的机制尚不清楚。我们推测，对受IRI影响的肾脏进行基因表达谱分析，将有助于明确IRI引发AR和CAN的机制。此外，我们还研究了当前免疫抑制药物的分子靶点如何受IRI影响。将C57BL/6J小鼠分为三组，分别接受30分钟（n = 3）或60分钟（n = 3）的双侧肾脏缺血处理，或进行假手术（n = 5）。在36小时时收集肾脏组织，使用Affymetrix 430MOEA（共22626个基因）芯片和GC-RMA-SAM流程进行分析。错误发现率（q < 1%）且变化倍数（FC）为+/-50%的基因被视为受IRI影响。利用基因本体论（GO）分析，筛选出编码组织相容性和抗原呈递因子、钙调神经磷酸酶以及雷帕霉素哺乳动物靶点（mTOR）通路相关蛋白的基因。GO分析确定，分别有10个和17个与同种免疫相关的基因受30分钟和60分钟缺血诱导的IRI影响，包括Traf6（FC = 2.99）和H2-D1（FC = 2.58）。我们还在分别由Fkbp5（FC = 4.18）和Fkbp1a（FC = 2.0）以及Eif4ebp1（FC = 16.8）和Akt1（FC = 3.64）代表的钙调神经磷酸酶和mTOR通路中检测到显著的IRI基因组反应。这些数据表明，IRI上调了多个与同种免疫相关的基因的表达，进而可能增强同种免疫反应。我们发现IRI诱导与钙调神经磷酸酶和mTOR通路相关的基因上调，这与临床观察结果一致，即FK506和雷帕霉素可以改变DGF的病程。对这些通路进行进一步验证和剖析，可能会带来新的方法，通过改善对早期“非免疫”移植事件的管理，降低对更典型的“免疫”变化和CAN的易感性。
