Proteomic analysis of saliva from patients with oral chronic graft-versus-host disease.
Chronic graft-versus-host disease (cGVHD) is an immune-mediated disorder and is the major long-term complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The oral mucosa, including the salivary glands, is affected in the majority of patients with cGVHD; however, at present there is only a limited understanding of disease pathobiology. In this study, we performed a quantitative proteomic analysis of saliva pooled from patients with and without oral cGVHD-cGVHD(+) and cGVHD(-), respectively-using isobaric tags for relative and absolute quantification labeling, followed by tandem mass spectrometry. Among 249 salivary proteins identified by tandem mass spectrometry, 82 exhibited altered expression in the oral cGVHD(+) group compared with the cGVHD(-) group. Many of the identified proteins function in innate or acquired immunity, or are associated with tissue maintenance functions, such as proteolysis or the cytoskeleton. Using ELISA immunoassays, we further confirmed that 2 of these proteins, IL-1 receptor antagonist and cystatin B, showed decreased expression in patients with active oral cGVHD (P < .003). Receiver operating curve characteristic analysis revealed that these 2 markers were able to distinguish oral cGVHD with a sensitivity of 85% and specificity of 60%, and showed slightly better discrimination in newly diagnosed patients evaluated within 12 months of allo-HSCT (sensitivity, 92%; specificity 73%). In addition to identifying novel potential salivary cGVHD biomarkers, our study demonstrates that there is coordinated regulation of protein families involved in inflammation, antimicrobial defense, and tissue protection in oral cGVHD that also may reflect changes in salivary gland function and damage to the oral mucosa.
Department of Pathology, University of Washington, Seattle, Washington.
Ivana
I
Devic
Department of Pathology, University of Washington, Seattle, Washington.
Min
M
Shi
Department of Oral Medicine, University of Washington, Seattle, Washington; Seattle Cancer Care Alliance, Seattle, Washington; Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington.
Mark M
MM
Schubert
Department of Oral Medicine, University of Washington, Seattle, Washington; Seattle Cancer Care Alliance, Seattle, Washington.
Michele
M
Lloid
Department of Oral Health Sciences, University of Washington, Seattle, Washington.
Kenneth T
KT
Izutsu
Department of Pathology, University of Washington, Seattle, Washington.
Catherine
C
Pan
Department of Oral Health Sciences, University of Washington, Seattle, Washington.
Melody
M
Missaghi
Department of Oral Health Sciences, University of Washington, Seattle, Washington.
Thomas H
TH
Morton
Department of Oral Health Sciences, University of Washington, Seattle, Washington.
Lloyd A
LA
Mancl
Department of Pathology, University of Washington, Seattle, Washington.
Jing
J
Zhang
Department of Oral Health Sciences, University of Washington, Seattle, Washington; Division of Dermatology, Department of Medicine, University of Washington, Seattle, Washington. Electronic address: rp@uw.edu.
Richard B
RB
Presland
D000328:Adult; D000368:Aged; D002908:Chronic Disease; D005260:Female; D006086:Graft vs Host Disease / Q000378:metabolism*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D009059:Mouth Diseases / Q000378:metabolism*; D009061:Mouth Mucosa / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012463:Saliva / Q000737:chemistry / Q000378:metabolism*
Biomarker discovery; Chronic graft-versus-host disease; Proteomics; Saliva; iTRAQ mass spectrometry