Objective    Non-invasive detection is the focus of intense research in diagnosis of acute renal allograft rejection currently. Urine protein is considered the cue to reflect the pathological changes in kidney disease. In this study

we explored the urine markers for early acute renal allograft rejection.     Methods    The urine protein of two patients with acute renal allograft rejection were examined by 2D gel electrophoresis and bioinformatics. We adopted pH 4-7 ready strip IPG and stained the gel with Sypro-Ruby. The digitized 2D maps of urine protein were quantitatively analyzed using 2D-analysis software packages. By analyzing the differential expressions of proteome between different time points (1

2

3 days before acute rejection and 7

14

21 days after acute rejection)

30 protein spots were selected and analyzed by MALDI-TOF-MS/MS.     Results    We obtained 2D gel electrophoresis maps of urine protein of the patients with acute renal allograft rejection

which are of good reproducibility and resolution. Sixteen protein spots were identified

resulting in thirteen corresponding proteins. Out of these proteins

we screened three proteins (alpha-1-antichymotrypsin

tumor rejection antigen gp96

Zn-Alpha-2-Glycoprotein) closely related to acute rejection.     Conclusion    The urine protein spots on 2D gel electrophoresis maps for the patients with acute renal allograft rejection were of obvious difference when detected at different time points of acute rejection. Alpha-1-antichymotrypsin

tumor rejection antigen gp96 and Zn-Alpha-2-Glycoprotein might be the candidate protein markers to diagnose acute renal allograft rejection after renal transplantation.