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.

Comparison of Two ELISA Methods and Mass Spectrometry for Measurement of Vitamin D-Binding Protein: Implications for the Assessment of Bioavailable Vitamin D Concentrations Across Genotypes

“…As detailed previously, (41) genotyping in CRIC was performed using the HumanCVD BeadChip V2 IBC ITMAT/Broad/CARE (IBC) Array (Illumina, Inc., San Diego, CA, USA), a gene-centric SNP array that includes approximately 50,000 SNPs in approximately 2100 candidate genes, at the Children's Hospital of Philadelphia Center for Applied Genomics. Samples from the CRIC study were excluded if: sample call rate was <0.97; there was reduced or excess heterozygosity (Inbreeding | F | <0.2); or there was evidence of cryptic relatedness (PI_HAT identity-by-descent <0.2).…”

Differentially Expressed RNA from Public Microarray Data Identifies Serum Protein Biomarkers for Cross-Organ Transplant Rejection and Other Conditions

“…For example, in renal transplant rejection, significantly increased protein concentrations of VEGF have been observed in serum and urine [21], [22], [23]. Increases in the following other entities have also been observed: CXCL9 in urine [24], soluble CD44 in plasma [25], ADL in serum [26], and TNF-alpha in serum [27]. HLA class I (ABC) protein levels were recently found to be elevated on the surfaces of peripheral blood CD3+/CD8+ T lymphocytes in AR at 14 and 21 days after renal transplantation [28].…”

Proteomic analysis of differentially expressed proteins in the serum of patients with acute renal allograft rejection using iTRAQ labelling technology

“…1,25-dihydroxycholecalciferol [1,25-(OH) 2 D 3 ; also known as calcitriol] is one of the active forms of vitamin D in the kidney. Previous studies have revealed that 1,25-(OH) 2 D 3 has an essential role in immunoregulation and allograft rejection ( 55 , 56 ), and VDBP may be one of the serum biomarker candidates of acute renal allograft rejection. These candidate proteins may provide a scientific foundation for the pathogenic mechanisms and potential therapeutic approach for AR that warrant further research.…”