Implication of granulocyte-macrophage colony-stimulating factor induced neutrophil gelatinase-associated lipocalin in pathogenesis of rheumatoid arthritis revealed by proteome analysis

In rheumatoid arthritis (RA), synovial fluid (SF)contains a large number of neutrophils that contribute to the inflammation and destruction of the joints. The SF also contains granulocyte-macrophage colony-stimulating factor (GM-CSF),which sustains viability of neutrophils and activates their functi...

Full description

Saved in:
Bibliographic Details
Published in:Arthritis research & therapy 2009-01, Vol.11 (1), p.R3-R3
Main Authors: Katano, Masayoshi, Okamoto, Kazuki, Arito, Mitsumi, Kawakami, Yuki, Kurokawa, Manae S, Suematsu, Naoya, Shimada, Sonoko, Nakamura, Hiroshi, Xiang, Yang, Masuko, Kayo, Nishioka, Kusuki, Yudoh, Kazuo, Kato, Tomohiro
Format: Article
Language:English
Subjects:
Acute-Phase Proteins - metabolism
Adenosine Triphosphatases - metabolism
Arthritis, Rheumatoid - metabolism
Cathepsin D - metabolism
Causes of
Cells, Cultured
Development and progression
Diagnosis
Granulocyte-macrophage colony stimulating factor
Granulocyte-Macrophage Colony-Stimulating Factor - physiology
GTP-Binding Proteins
Health aspects
Humans
Lipocalin-2
Lipocalins - metabolism
Neutrophils
Neutrophils - metabolism
Physiological aspects
Proteomics
Proto-Oncogene Proteins - metabolism
Rheumatoid arthritis
Transglutaminases - metabolism
Up-Regulation
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In rheumatoid arthritis (RA), synovial fluid (SF)contains a large number of neutrophils that contribute to the inflammation and destruction of the joints. The SF also contains granulocyte-macrophage colony-stimulating factor (GM-CSF),which sustains viability of neutrophils and activates their functions. Using proteomic surveillance, we here tried to elucidate the effects of GM-CSF on neutrophils. Neutrophils stimulated by GM-CSF were divided in to four subcellular fractions: cytosol, membrane/organelle, nuclei,and cytoskeleton. Then, proteins were extracted from each fraction and digested by trypsin. The produced peptides were detected using matrix-assisted laser desorption ionisation-time of-flight mass spectrometry (MALDI-TOF MS). We detected 33 peptide peaks whose expression was upregulated by more than 2.5-fold in GM-CSF stimulated neutrophils and identified 11 proteins out of the 33 peptides using MALDI-TOF/TOF MS analysis and protein database searches. One of the identified proteins was neutrophil gelatinase-associated lipocalin (NGAL). We confirmed that the level of NGAL in SF was significantly higher in patients with RA than in those with osteoarthritis. We next addressed possible roles of the increased NGAL in RA. We analysed proteome alteration of synoviocytes from patients with RA by treatment with NGAL in vitro. We found that, out of the detected protein spots (approximately 3,600 protein spots), the intensity of 21 protein spots increased by more than 1.5-fold and the intensity of 10 protein spots decreased by less than 1 to 1.5-fold as a result of the NGAL treatment. Among the 21 increased protein spots, we identified 9 proteins including transitional endoplasmic reticulum ATPase (TERA), cathepsin D, and transglutaminase 2 (TG2), which increased to 4.8-fold, 1.5-fold and 1.6-fold, respectively. Two-dimensional electrophoresis followed by western blot analysis confirmed the upregulation of TERA by the NGAL treatment and, moreover, the western blot analysis showed that the NGAL treatment changed the protein spots caused by post-translational modification of TERA.Furthermore, NGAL cancelled out the proliferative effects of fibroblast growth factor (FGF)-2 and epidermal growth factor(EGF) on chondrocytes from a patient with RA and proliferative effect of FGF-2 on chondrosarcoma cells.Conclusions Our results indicate that GM-CSF contributes to the pathogenesis of RA through upregulation of NGAL in neutrophils, followed by induction of TERA, cathep
ISSN:1478-6354
1478-6362
DOI:10.1186/ar2587