基因差异显示法在器官移植排斥反应预测中的应用  

Application DDRT-PCR in transplant rejection study

作　　者：李文[1] 周健[2] 刘艳霞[1] 王夏[2] 刘俊敏[3] 薛健 孟旭[4] 黄益民[1] 

机构地区：[1]北京市心肺血管疾病研究所细胞免疫室 [2]中国科学院微生物研究所 [3]北京安贞医院泌尿外科 [4]北京安贞医院心外科

出　　处：《中华微生物学和免疫学杂志》2002年第2期148-151,共4页Chinese Journal of Microbiology and Immunology

基　　金：北京市卫生局重点学科基金资助项目;北京市科学技术委员会重点实验室研究基金资助项目

摘　　要：目的　用基因差异显示 (DDRT PCR)方法观察器官移植后患者白细胞mRNA表达变化 ,为建立无创性预测免疫排斥反应指征提供依据。方法　对 1例心脏移植和 1例心脏瓣膜置换术加肾移植患者术后外周血白细胞mRNA表达进行DDRT PCR分析 ,并对差异条带进行测序。结果　差异分析显示 ,(1)在肾移植术中开放动脉后 ,即有Rab oncogene、SLAM (signalinglymphocyticactivationmolecule)、酪氨酸蛋白激酶P5 9fyn、IAP(inhibitsapoptosisprotein) family等基因的表达 ,于术后 72h～ 5d降低 ,术后 7d表达出现高峰 ;在术后 5d有ZincfingerA基因表达。 (2 )心脏移植术后 10d ,有 10条cDNA条带的灰度发生了不规则变化 ,它们分别为核糖体蛋白、鲨烯合成酶、信号识别颗粒、mRNA识别蛋白、酪氨酸激酶和FK5 0 6结合蛋白等基因。结论　本实验所得的异常基因表达比临床监测指标变化提前 3～ 5d。应用DDRTs ] Objective To observe the quantity change of leucocytes mRNA expression after organ transplant by using DDRT-PCR(mRNA differential display reverse transcription PCR) in order to detect immun rejection, and develop a no-injured monitor methods. Methods Using DDRT-PCR method to analyze leucocytes mRNA on a heart transplant patient and an AVR+MVR+kidney transplant patients, sequenced the differential samples. Results 1. Kidney transplant patient, the genes of Rab-oncogene, SLAM, P59 fyn, IAP-family are expressed when kidney artery opened again and all then decreased 72 h-5 d after operation, then increased and got highest after 7 d, ZincfingerA is expressed 5 d after operation. 2. In heart transplant patient, DDRT-PCR found that 10 cDNA have irregular change after operation, the gene include: RPL23, RPL24, SRP68, REA1, P59 fyn, FKBP1A, et al. Conclusion Differential gene expression was found with some changes 3-5 d, earlier than the clinical monitor test results. Using DDRT-PCR methods, we could observe activating process of leucocytes, recognized the regular change of multiple genes which is related to activating leucocytes in early phase of organ transplantation.

关 键 词：器官移植 基因表达 差异显示分析 免疫排斥 外周血白细胞 预测 

分 类 号：R392.4[医药卫生—免疫学]