Two-dimensional difference gel electrophoresis urinary proteomic profile in the search of nonimmune chronic allograft dysfunction biomarkers.
BACKGROUND
Despite advances in therapeutics, graft loss associated with chronic allograft dysfunction (CAD) remains high. Urinary proteomic analysis is a noninvasive method that could be used to detect and evaluate CAD in renal transplant recipients. This study was aimed to establish the normal proteome map of stable transplant patients and to validate the utility of two-dimensional difference gel electrophoresis (2DE-DIGE) in identifying new candidates as urinary biomarkers of CAD.


METHODS
Morning spot urine samples that were collected from kidney transplant recipients with biopsy-proven interstitial fibrosis and tubular atrophy (IFTA) stages 0-I-II/III (n=8/group) under immunosuppressive treatment with tacrolimus plus mycophenolate with or without prednisone. 2DE silver staining and mass spectrometry analyses were used to establish the normal proteome map, and 2DE-DIGE and mass spectrometry were used to identify proteins exhibiting differential abundance.


CONCLUSIONS
This study defines the normal proteome of stable renal transplant patients, which is composed of several plasma proteins, as well as of immunologic proteins that are probably specific to transplant recipients. The 2DE-DIGE study showed 19 proteins with differential concentrations, depending on the IFTA histologic score. These 19 proteins could be used as urinary biomarkers of the severity of IFTA in renal transplant recipients.
Laboratori Experimental de Nefrologia i Trasplantament Renal (LENIT), Departament de Nefrologia I Trasplantament Renal, Hospital Clínic, IDIBAPS, Universitat de Barcelona, Barcelona, Spain. ebanon@clinic.ub.es
Elisenda
E
Bañón-Maneus
Fritz
F
Diekmann
Montserrat
M
Carrascal
Luis F
LF
Quintana
Daniel
D
Moya-Rull
Monica
M
Bescós
Maria J
MJ
Ramírez-Bajo
Jordi
J
Rovira
Alex
A
Gutierrez-Dalmau
Amanda
A
Solé-González
Joaquin
J
Abián
Josep M
JM
Campistol
D000595:Amino Acid Sequence; D001284:Atrophy; D015415:Biomarkers / Q000652:urine; D001706:Biopsy; D004359:Drug Therapy, Combination; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D045604:Extracellular Fluid; D006801:Humans; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D007525:Isoelectric Focusing / Q000379:methods; D016030:Kidney Transplantation / Q000276:immunology / Q000473:pathology*; D007684:Kidney Tubules / Q000473:pathology; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D011183:Postoperative Complications / Q000473:pathology; D011507:Proteinuria / Q000235:genetics* / Q000378:metabolism; D020543:Proteome / Q000235:genetics*; D014184:Transplantation, Homologous / Q000473:pathology*
D015415:Biomarkers; D007166:Immunosuppressive Agents; D010455:Peptides; D020543:Proteome