Quantitative Proteomic Analysis of Peripheral Blood Mononuclear Cells in Ankylosing Spondylitis by iTRAQ

Clin Transl Sci. 2015 Oct;8(5):579-83. doi: 10.1111/cts.12265. Epub 2015 Mar 19.

Abstract

This study was designed to identify and quantify the different proteins expression levels in ankylosing spondylitis (AS) and to explore the pathogenesis of AS. We performed isobaric tags for relative and absolute quantitation (iTRAQ) coupled with multiple chromatographic fractionation and tandem mass spectrometry to detect the proteins profiling in peripheral blood mononuclear cells (PBMCs) from AS patients and healthy controls. Mascot software and the International Protein Index and the Gene Ontology (GO) database were used to conduct the bioinformatics analysis. The differentially expressed proteins were validated by enzyme-linked immunosorbent assay (ELISA). A total of 1,232 proteins were identified by iTRAQ, of which 183 showed differential expression and 18 differentially expressed proteins were acute phase reactants. Upon mapping of the differentially expressed proteins to GO database, we found four differentially expressed proteins involved in the biological process of cell killing, including up-regulated cathepsin G (CTSG), neutrophil defensin3 (DEFA3), protein tyrosine phosphatase receptor type C (PTPRC), and down-regulated peroxiredoxin-1(PRDX1),which were consistent with the verified results of ELISA. Our proteomic analyses suggested that the proteins involved in the biological process of cell killing might play an important role in the pathogenesis of AS.

Keywords: ankylosing spondylitis; cell killing; iTRAQ; proteomic.

MeSH terms

  • Adult
  • Biomarkers / blood
  • Blood Proteins / metabolism*
  • Case-Control Studies
  • Cathepsin G / blood
  • Chromatography, High Pressure Liquid
  • Computational Biology
  • Databases, Protein
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Leukocyte Common Antigens / blood
  • Leukocytes, Mononuclear / metabolism*
  • Male
  • Middle Aged
  • Peroxiredoxins / blood
  • Proteomics / methods*
  • Reproducibility of Results
  • Spondylitis, Ankylosing / blood*
  • Tandem Mass Spectrometry
  • Young Adult
  • alpha-Defensins / blood

Substances

  • Biomarkers
  • Blood Proteins
  • alpha-Defensins
  • human neutrophil peptide 3
  • PRDX1 protein, human
  • Peroxiredoxins
  • Leukocyte Common Antigens
  • PTPRC protein, human
  • CTSG protein, human
  • Cathepsin G