肾移植患者尿液中供者细胞DNA变化在急性排斥反应诊断中的意义

Diagnosis role of urinary donor-derived DNA in acute renal allograft rejection

目的 :探讨肾移植受者尿液中供者细胞的出现与急性排斥反应的相关关系及其临床意义。方法 :以供者为男性、受者为女性或HLA DR抗原有错配的 80例肾移植受者为研究对象 ,定期收集尿液标本 ,从中提取DNA ,利用特异性引物聚合酶链反应分别检测Y染色体上特异的基因片段DYZ 1和HLA DR抗原的基因DRB 1。结果 :手术当天受者的尿液中即有供者细胞出现 ,随着时间的推移 ,尿液中供者细胞DNA的基因表达强度逐渐减弱 ,直至术后 30天 ,仍有 90 0 %受者的尿液中有供者细胞DNA的基因表达 ,其中 8例 ( 2 9 6 % )发生了急性排斥反应 ;出院后发生急性排斥反应者 ,90 0 %的尿液标本中能检测到供者细胞DNA ,抗排斥治疗结束后 2周 ,83 3%转为阴性 ;肾功能良好的稳定期受者 ,仅 6 7%的受者尿液中DYZ 1或HLA DRB1基因阳性。结论 :肾移植受者尿中供者细胞DNA的检测可以作为诊断急性排斥反应的一种方法 。

Objective:To investigate the relationship between the appearance of donor cells in urine and acute rejection and the clinical implication.Methods:Eighty renal transplantation patients were observed,in which the donors were male and the recipients were female,or HLA-DR antigen were mismatched (30 cases were at perioperative period,20 cases were subjected to acute rejection,30 cases had stable renal function).Urine samples were collected regularly.PCR and PCR-SSP were applied to detect DYZ-1 (special gene fragment of Y-chromosome) and DRB-1 of HLA-DR antigen respectively after DNA were obtained.Results:Perioperative period group:donor cells in urine were detected in all the patients 24 h after operation.With the development of disease,the intensity of donor DNA expression in urine was decreased generally.30 days later,donor cells in urine disappeared only in 3 cases of 30 cases,and acute rejection happened in 8 cases of the rest 27 cases.Acute rejection group:donor cells in urine were detected in 18 cases(90%);2 weeks following anti-rejection therapy,donor cells in urine were negative only in 3 cases,still positive in the other 15 cases,and the intensity of donor DNA expression in urine was decreased generally during the treatment.Donor cells in urine were negative in 16 cases(88 9%) after treatment for 3 months.Stable renal function group:DYZ-1 or HLA-DRB1 was positive in 2 cases(6 7%),negative in 28 cases(93 3%).Conclusion:PCR and PCR-SSP were used to detect DNA of donor cells in urine,which would be a new method to diagnose acute rejection of renal transplantation,but would not exactly fit for those happened in early stage.The intensity change of donor DNA expression in urine represented the recovery of renal transplantation,which provided the possibility to evaluate renal allograft rejection quantitatively at the same time.