Identification of parotid salivary biomarkers in Sjogren's syndrome by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry and two-dimensional difference gel electrophoresis

Rheumatology (Oxford). 2006 Sep;45(9):1077-86. doi: 10.1093/rheumatology/kei212. Epub 2006 Mar 7.

Abstract

Objectives: To identify the most significant salivary biomarkers in Sjögren's syndrome (SS) using proteomic methods.

Methods: Parotid saliva from 20 non-SS subjects and 41 primary SS patients was analysed. Protein expression profiles for each sample were generated by surface-enhanced laser desorption/ionization time-of-flight-mass spectrometry (SELDI-TOF-MS). Mean peak intensities of SS patients and non-SS subjects were compared by univariate analyses. Samples pooled by diagnosis (SS and non-SS) and labelled with different Cy dyes were compared by two-dimensional difference gel electrophoresis (2D-DIGE). Two protein levels that were most significantly different by SELDI-TOF-MS and 2D-DIGE were validated by enzyme-linked immunosorbent assay in individual samples.

Results: SELDI-TOF-MS of 10-200 kDa peaks revealed eight peaks with >2-fold changes in the SS group that differed from non-SS at P < 0.005. Peaks of 11.8, 12.0, 14.3, 80.6 and 83.7 kDa were increased, while 17.3, 25.4, and 35.4 kDa peaks were decreased in SS samples. 2D-DIGE identified significant increases of beta-2-microglobulin, lactoferrin, immunoglobulin (Ig) kappa light chain, polymeric Ig receptor, lysozyme C and cystatin C in all stages of SS. Two presumed proline-rich proteins, amylase and carbonic anhydrase VI, were reduced in the patient group. Three of these ten biomarkers have not been associated previously with SS.

Conclusions: The salivary proteomic profile of SS is a mixture of increased inflammatory proteins and decreased acinar proteins when compared with non-SS. Future studies will test the ability of these biomarker levels, alone and in combination, to diagnose the salivary component of SS.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Amylases / analysis
  • Biomarkers / analysis
  • Carbonic Anhydrases / analysis
  • Case-Control Studies
  • Cystatin C
  • Cystatins / analysis
  • Electrophoresis, Gel, Two-Dimensional / methods
  • Humans
  • Lactoferrin / analysis*
  • Muramidase / analysis
  • Parotid Gland*
  • Protein Subunits / analysis
  • Proteomics
  • Receptors, Polymeric Immunoglobulin / analysis
  • Saliva / chemistry*
  • Sjogren's Syndrome / diagnosis*
  • Sjogren's Syndrome / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Statistics, Nonparametric
  • beta 2-Microglobulin / analysis*

Substances

  • Biomarkers
  • CST3 protein, human
  • Cystatin C
  • Cystatins
  • Protein Subunits
  • Receptors, Polymeric Immunoglobulin
  • beta 2-Microglobulin
  • Amylases
  • Muramidase
  • lysozyme C, human
  • Lactoferrin
  • Carbonic Anhydrases
  • carbonic anhydrase VI