Journal of Clinical Immunology

, Volume 32, Issue 6, pp 1390–1399

Quantitative Salivary Proteomic Differences in Oral Chronic Graft-versus-Host Disease

  • Carol W Bassim
  • Kiran S. Ambatipudi
  • Jacqueline W. Mays
  • Dean A. Edwards
  • Stephan Swatkoski
  • Helen Fassil
  • Kristin Baird
  • Marjan Gucek
  • James E. Melvin
  • Steven Z. Pavletic
Article

Abstract

Purpose

Chronic graft-versus-host disease (cGVHD) is a severe immunological complication that occurs after allogeneic hematopoietic stem cell transplantation (HSCT). Although oral cGVHD occurs in >25 % of cGVHD patients and leads to decreased quality of life, its etiology is poorly understood. The present retrospective cross-sectional analysis of oral cGVHD patients sought to (1) test the feasibility of liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify protein biomarkers of oral cGVHD and (2) to gain a clearer understanding of salivary proteins impacted by oral cGVHD.

Methods

Using unstimulated whole saliva, we compared pooled saliva from five patients with a diagnosis of moderate or severe oral cGVHD, with a gender-and age- matched pool of five cGVHD patients with no oral mucosal findings. LC-MS/MS was used to identify salivary proteins, followed by Ingenuity Pathway Analysis (IPA). Selected mass spectrometric findings, including lactotransferrin, lactoperoxidase, and albumin, were confirmed by targeted label-free quantification.

Results

LC-MS/MS led to confident identification of 180 proteins. Of these proteins, 102 changed in abundance at least 2 fold, including 12 proteins identified only in the No oral cGVHD group. Downregulation of ~0.4 fold was confirmed for both lactotransferrin and lactoperoxidase in Oral cGVHD saliva using targeted label-free quantification. IPA analysis implicated pathways involved in cellular metabolism and immunoregulation.

Conclusions

Reduction of salivary lactoperoxidase, lactotransferrin, and several cysteine proteinase inhibitor family proteins suggests impaired oral antimicrobial host immunity in cGVHD patients. This shotgun proteomic analysis of oral cGVHD saliva using targeted label-free quantification of select proteins supports the use of mass spectrometry for future validation in a large patient population as noninvasive tests for screening, early detection, and monitoring of cGVHD.

Keywords

Saliva oral GVHD biomarker proteomics label-free quantitation transplant graft versus host 

Supplementary material

10875_2012_9738_MOESM1_ESM.xlsx (23 kb)
Supplementary Table 1Differentially expressed proteins identified in whole saliva with a minimum of two unique peptides in No oral cGVHD and Oral cGVHD groups. (XLSX 22 kb)

Copyright information

© Springer Science+Business Media, LLC (outside the USA) 2012

Authors and Affiliations

  • Carol W Bassim
    • 1
  • Kiran S. Ambatipudi
    • 1
  • Jacqueline W. Mays
    • 1
  • Dean A. Edwards
    • 1
  • Stephan Swatkoski
    • 3
  • Helen Fassil
    • 2
  • Kristin Baird
    • 2
  • Marjan Gucek
    • 3
  • James E. Melvin
    • 1
  • Steven Z. Pavletic
    • 2
  1. 1.National Institute of Dental and Craniofacial ResearchBethesdaUSA
  2. 2.National Cancer InstituteBethesdaUSA
  3. 3.Proteomics Core Facility, National Institute of Heart, Lung and Blood Institute, National Institutes of HealthBethesdaUSA

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