Characterization of acute renal allograft rejection by human serum proteomic analysis

Abstract

To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS) were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups: acute rejection (AR), stable renal function (SRF) and normal volunteer (N). Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.

Keywords

acute rejection / two-dimensional differential in-gel electrophoresis / reversed phase high-performance liquid chromatography / electrospray ionization mass spectrometry / ELISA / serum

References

[1] HariharanS., JohnsonC.P., BresnahanB.A., et al.. Improved graft survival after renal transplantation in the United States, 1998–1996. N Engl J Med, 2000, 342(9): 605-612

[2] McLarenA.J., FuggleS.V., WelshK.I., et al.. Chronic allograft failure in human renal transplantation: a multivariate risk factor analysis. Ann Surg, 2000, 232(1): 98-103

[3] BenfieldM.R., HerrinJ., FeldL., et al.. Safety of kidney biopsy in pediatric transplantation: a report of the controlled clinical trials in pediatric transplantation trial of induction therapy study group. Transplantation, 1999, 67(4): 544-547

[4] TraumA.Z., SchachterA.D.. Transplantation proteomics. Pediatr Transplant, 2005, 9(6): 700-711

[5] SolezK., ColvinR.B., RacusenL.C., et al.. Banff’ 05 Meeting Report: differential diagnosis of chronic allograft injury and elimination of chronic allograft nephropathy (’CAN’). Am J Transplant, 2007, 7(3): 518-526

[6] PengJ., EliasJ.E., ThoreenC.C., et al.. Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for large-scale protein analysis: the yeast proteome. J Proteome Res, 2003, 2(1): 43-50

[7] AndersonN.L., PolanskiM., PieperR., et al.. The human plasma proteome: a nonredundant list developed by combination of four separate sources. Mol Cell Proteomics, 2004, 3(4): 311-326

[8] HuangH.L., StasykT., MorandellS., et al.. Biomarker discovery in breast cancer serum using 2-D differential gel electrophoresis/ MALDI-TOF/TOF and data validation by routine clinical assays. Electrophoresis, 2006, 27(8): 1641-1650

[9] SchaubS., WilkinsJ.A., NickersonP.. Proteomics and renal transplantation: searching for novel biomarkers and therapeutic targets. Contrib Nephrol, 2008, 160: 65-75

[10] ClarkeW.. Proteomic research in renal transplantation. Ther Drug Monit, 2006, 28(1): 19-22

[11] SchaubS., WilkinsJ.A., AntonoviciM., et al.. Proteomic-based identification of cleaved urinarybeta2-microglobulin as a potential marker for acute tubular injury in renal allografts. Am J Transplant, 2005, 5(4Pt1): 729-738

[12] WittkeS., HaubitzM., WaldenM., et al.. Detection of acute tubulointerstitial rejection by proteomic analysis of urinary samples in renal transplant recipients. Am J Transplant, 2005, 5(10): 2479-2488

[13] El EssawyB., OtuH.H., ChoyB., et al.. Proteomic analysis of the allograft response. Transplantation, 2006, 82(2): 267-274

[14] VosholH., BrendlenN., MüllerD., et al.. Evaluation of biomarker discovery approaches to detect protein biomarkers of acute renal allograft rejection. J Proteome Res, 2005, 4(4): 1192-1199

[15] YuK.H., RustgiA.K., BlairI.A.. Characterization of proteins in human pancreatic cancer serum using differential gel electrophoresis and tandem mass spectrometry. J Proteome Res, 2005, 4(5): 1742-1751

[16] SchiødtF.V., RossaroL., StravitzR.T., et al.. Gc-globulin and prognosis in acute liver failure. Liver Transpl, 2005, 11(10): 1223-1227

[17] SpeeckaertM.M., GlorieuxG.L., VanholderR., et al.. Vitamin D binding protein and the need for vitamin D in hemodialysis patients. J Ren Nutr, 2008, 18(5): 400-407

[18] ZellaL.A., ShevdeN.K., HollisB.W., et al.. Vitamin D-binding protein influences total circulating levels of 1,25-dihydroxyvitamin D3 but does not directly modulate the bioactive levels of the hormone in vivo. Endocrinology, 2008, 149(7): 3656-3667

[19] ChristakosS., DhawanP., LiuY., et al.. New insights into the mechanisms of vitamin D action. J Cell Biochem, 2003, 88(4): 695-705

[20] MathieuC., JafariM.. Immunomodulation by 1,25-dihydroxyvitamin D3: therapeutic implications in hemodialysis and renal transplantation. Clin Nephrol, 2006, 66(4): 275-283

[21] KogaY., NaraparajuV.R., YamamotoN.. Antitumor effect of vitamin D-binding protein-derived macrophage activating factor on Ehrlich ascites tumor-bearing mice. Proc Soc Exp Biol Med, 1999, 220(1): 20-26

[22] YamamotoN., SuyamaH., UshijimaN.. Immunotherapy of metastatic breast cancer patients with vitamin D-binding protein-derived macrophage activating factor (GcMAF). Int J Cancer, 2008, 122(2): 461-467

[23] YamamotoN., SuyamaH.. Immunotherapy for prostate cancer with gc protein-derived macrophage-activating factor, GcMAF. Transl Oncol, 2008, 1(2): 65-72

[24] NickeleitV., AndreoniK.. Inflammatory cells in renal allografts. Front Biosci, 2008, 13: 6202-6213

[25] QiF., AdairA., FerenbachD., et al.. Depletion of cells of monocyte lineage prevents loss of renal microvasculature in murine kidney transplantation. Transplantation, 2008, 86(9): 1267-1274

This project was supported by a grant from National Basic Research 973 Program of China (No. 2009CB522407).