Yamayoshi Yasuko, Watanabe Toshihiko, Tanabe Minoru, Hoshino Ken, Matsumoto Koshi, Morikawa Yasuhide, Shimadzu Motohide, Kitajima Masaki, Tanigawara Yusuke
Department of Hospital Pharmacy, School of Medicine, Keio University, Tokyo, Japan.
Transplantation. 2006 Aug 15;82(3):320-6. doi: 10.1097/01.tp.0000228909.49640.08.
Because no biomarker that reflects small bowel allograft rejection is available, we applied surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) to develop noninvasive markers required for routine diagnosis.
Heterotopic small bowel transplantation (SBT) was performed in rats, and they were divided into four experimental groups: sham-operated rats (sham), syngeneic transplants (syngeneic), allogeneic transplants (allogeneic), allogeneic transplants received FK506 (allo+FK). Plasma samples were analyzed with SELDI ProteinChip arrays to detect peaks that predominated in the allogeneic model. Possible biomarkers were identified in combination with SELDI retentate chromatography mass spectrometry (RCMS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The identified protein was further analyzed by immunohistochemistry.
An increase in the level of a 14.8-kDa protein, identified as lysozyme, was observed specifically in the plasma of the allogeneic group; the levels of this protein remained unchanged in the plasma of the other groups. On the other hand, the levels of a 10.1-kDa and a 13.0-kDa protein, identified as migration inhibitory factor-related proteins (MRP), MRP-8 and MRP-14, respectively, began to increase from an early stage of acute rejection. We also observed that lysozyme-positive macrophages had strongly infiltrated the lamina propria during acute rejection.
We identified three plasma proteins-MRP-8, MRP-14, and lysozyme-that increased during small bowel allograft rejection. The identified proteins appeared to be markers for inflammation associated with allograft rejection. This proteomic approach will be useful for the identification of candidate biomarkers.
由于目前尚无能够反映小肠移植排斥反应的生物标志物，我们应用表面增强激光解吸/电离飞行时间质谱（SELDI-TOF MS）技术来开发常规诊断所需的非侵入性标志物。
对大鼠进行异位小肠移植（SBT），并将其分为四个实验组：假手术大鼠（假手术组）、同基因移植组（同基因组）、异基因移植组（异基因组）、接受FK506的异基因移植组（异基因+FK组）。采用SELDI蛋白芯片阵列分析血浆样本，以检测在异基因模型中占主导地位的峰。结合SELDI保留物色谱质谱（RCMS）和基质辅助激光解吸/电离质谱（MALDI-MS）鉴定可能的生物标志物。对鉴定出的蛋白质进一步进行免疫组织化学分析。
在异基因组血浆中特异性观察到一种14.8 kDa蛋白质（鉴定为溶菌酶）水平升高；该蛋白质在其他组血浆中的水平保持不变。另一方面，一种10.1 kDa蛋白质（鉴定为迁移抑制因子相关蛋白（MRP）-8）和一种13.0 kDa蛋白质（鉴定为MRP-14）的水平从急性排斥反应早期开始升高。我们还观察到在急性排斥反应期间，溶菌酶阳性巨噬细胞强烈浸润固有层。
我们鉴定出三种在小肠移植排斥反应期间水平升高的血浆蛋白——MRP-8、MRP-14和溶菌酶。鉴定出的蛋白质似乎是与移植排斥反应相关的炎症标志物。这种蛋白质组学方法将有助于鉴定候选生物标志物。
