Ankylosing spondylitis monocytes show upregulation of proteins involved in inflammation and the ubiquitin proteasome pathway

Objectives: To determine if peripheral blood monocytes from patients with ankylosing spondylitis (AS) differed in protein expression compared to rheumatoid arthritis (RA) and healthy controls (HC). Methods: Monocyte protein expression was characterised by 2D gel electrophoresis and by label-free quantitative expression profiling, using nano-ultra performance liquid chromatography coupled to electrospray ionisation mass spectrometry (ESI-MSE, where E refers to low/high collision energy switching). Data sets were analysed using the Waters expression profiling system and Ingenuity pathway analysis (IPA). Results: Two-dimensional gel electrophoresis showed upregulation of proteasomal constituents in AS monocytes, including the β subunit of proteasome activator (PA)28. Monocyte expression profiling and IPA showed that significant changes in protein expression within the ubiquitin proteasome pathway (UPP) were restricted to AS monocytes. Statistically significant differences in protein expression involving the leucocyte extravasation, vascular endothelial growth factor, integrin and Toll-like receptor signalling pathways were seen in AS and RA monocytes compared to healthy controls. No evidence of upregulation of proteins involved in the endoplasmic reticulum stress response pathway was found in either AS or RA monocytes. Finally, the PA28 complex was shown to increase the generation of human leucocyte antigen (HLA)-B27 antigenic epitopes by the proteasome in vitro. Conclusions: Our proteomic analyses support the hypothesis that monocytes play an important role in the pathogenesis of AS and RA, and further suggest a specific role in AS for the UPP. Quantitative proteomic expression profiling constitutes a powerful new tool for rheumatology research.

[1]  A. Görg,et al.  Improved silver staining protocols for high sensitivity protein identification using matrix‐assisted laser desorption/ionization‐time of flight analysis , 2001, Proteomics.

[2]  B. Ulfhake,et al.  Iron load and redox stress in skeletal muscle of aged rats , 2007, Muscle & nerve.

[3]  A. Lubben,et al.  Ultra-fast tandem mass spectrometry scanning combined with monolithic column liquid chromatography increases throughput in proteomic analysis. , 2006, Rapid communications in mass spectrometry : RCM.

[4]  J. Y. Park,et al.  Influence of VEGF gene polymorphisms on the severity of ankylosing spondylitis. , 2005, Rheumatology.

[5]  S. Mikkat,et al.  Proteomic analysis of peripheral blood mononuclear cells: selective protein processing observed in patients with rheumatoid arthritis. , 2007, Journal of proteome research.

[6]  W. MacNee,et al.  Potential role of IL‐8, platelet‐activating factor and TNF‐α in the sequestration of neutrophils in the lung: effects on neutrophil deformability, adhesion receptor expression, and chemotaxis , 2002, European journal of immunology.

[7]  R. Inman,et al.  Gene expression analysis of macrophages derived from ankylosing spondylitis patients reveals interferon-gamma dysregulation. , 2008, Arthritis and rheumatism.

[8]  K. Xu,et al.  IFN-gamma and TNF regulate macrophage expression of the chemotactic S100 protein S100A8. , 2000, Journal of immunology.

[9]  D. Nixon,et al.  Structural homologies between two HLA B27-restricted peptides suggest residues important for interaction with HLA B27. , 1990, International immunology.

[10]  A. Cats,et al.  Evaluation of diagnostic criteria for ankylosing spondylitis. A proposal for modification of the New York criteria. , 1984, Arthritis and rheumatism.

[11]  P. Kloetzel,et al.  Identification of HLA-B27-Restricted Peptides from the Chlamydia trachomatis Proteome with Possible Relevance to HLA-B27-Associated Diseases1 , 2001, The Journal of Immunology.

[12]  R. Colbert,et al.  HLA-B27 up-regulation causes accumulation of misfolded heavy chains and correlates with the magnitude of the unfolded protein response in transgenic rats: Implications for the pathogenesis of spondylarthritis-like disease. , 2007, Arthritis and rheumatism.

[13]  M. Xiong,et al.  A 588-gene microarray analysis of the peripheral blood mononuclear cells of spondyloarthropathy patients. , 2002, Rheumatology.

[14]  M. Blumenberg,et al.  Effects of tumor necrosis factor-alpha (TNF alpha) in epidermal keratinocytes revealed using global transcriptional profiling. , 2004, The Journal of biological chemistry.

[15]  T. Tsuda,et al.  Cytokine‐induced expression of an inducible type of nitric oxide synthase gene in cultured vascular smooth muscle cells , 1993, FEBS letters.

[16]  A. Osterhaus,et al.  Antigenic Drift in the Influenza A Virus (H3N2) Nucleoprotein and Escape from Recognition by Cytotoxic T Lymphocytes , 2000, Journal of Virology.

[17]  Michael J. Wilson,et al.  Cutting Edge: Leukocyte Receptor Complex-Encoded Immunomodulatory Receptors Show Differing Specificity for Alternative HLA-B27 Structures1 , 2001, The Journal of Immunology.

[18]  S. Chouaib,et al.  The induction of nitric oxide by interleukin-12 and tumor necrosis factor-alpha in human natural killer cells: relationship with the regulation of lytic activity. , 1998, Blood.

[19]  L. Ivashkiv,et al.  IFN-γ-Primed Macrophages Exhibit Increased CCR2-Dependent Migration and Altered IFN-γ Responses Mediated by Stat11 , 2005, The Journal of Immunology.

[20]  L. Ivashkiv,et al.  IFN-gamma-primed macrophages exhibit increased CCR2-dependent migration and altered IFN-gamma responses mediated by Stat1. , 2005, Journal of immunology.

[21]  K. Mossman.  The Wellcome Trust Case Control Consortium, U.K. , 2008 .

[22]  B. Han,et al.  TNFα-Induced Long Pentraxin PTX3 Expression in Human Lung Epithelial Cells via JNK , 2005, The Journal of Immunology.

[23]  Carolyn L. Geczy,et al.  IFN-γ and TNF Regulate Macrophage Expression of the Chemotactic S100 Protein S100A81 , 2000, The Journal of Immunology.

[24]  C. Greenwood,et al.  VEGF, FGF1, FGF2 and EGF gene polymorphisms and psoriatic arthritis , 2007, BMC musculoskeletal disorders.

[25]  M. Fink,et al.  Ethyl Pyruvate Ameliorates Intestinal Epithelial Barrier Dysfunction in Endotoxemic Mice and Immunostimulated Caco-2 Enterocytic Monolayers , 2003, Journal of Pharmacology and Experimental Therapeutics.

[26]  S. Westerheide,et al.  PTEN Blocks Tumor Necrosis Factor-induced NF-κB-dependent Transcription by Inhibiting the Transactivation Potential of the p65 Subunit* , 2002, The Journal of Biological Chemistry.

[27]  Paul Wordsworth,et al.  HLA-B associations of HLA-B27 negative ankylosing spondylitis: comment on the article by Yamaguchi et al. , 1996, Arthritis and rheumatism.

[28]  Hong Wang,et al.  Transforming Growth Factor-β1 Inhibits Cytokine-mediated Induction of Human Metalloelastase in Macrophages* , 2000, The Journal of Biological Chemistry.

[29]  M. Blumenberg,et al.  Effects of Tumor Necrosis Factor-α (TNFα) in Epidermal Keratinocytes Revealed Using Global Transcriptional Profiling* , 2004, Journal of Biological Chemistry.

[30]  E. Lightfoot,et al.  CD8αβ T Cells Are Not Essential to the Pathogenesis of Arthritis or Colitis in HLA-B27 Transgenic Rats1 , 2003, The Journal of Immunology.

[31]  A. Dey,et al.  Osmotic Response Element Is Required for the Induction of Aldose Reductase by Tumor Necrosis Factor-α* , 1999, The Journal of Biological Chemistry.

[32]  V. Shashoua,et al.  Protein release from hippocampus in vitro , 1984, Brain Research.

[33]  F. Arnett.  Revised criteria for the classification of rheumatoid arthritis. , 1990, Orthopedic nursing.

[34]  Hans-Georg Rammensee,et al.  A role for the proteasome regulator PA28α in antigen presentation , 1996, Nature.

[35]  E. Friauf,et al.  A subcellular prefractionation protocol for minute amounts of mammalian cell cultures and tissue , 2005, Proteomics.

[36]  H. Clevers,et al.  Proteome changes induced by knock-down of the deubiquitylating enzyme HAUSP/USP7. , 2007, Journal of proteome research.

[37]  C. Lindgren,et al.  •Australo-Anglo-American Spondylitis Consortium (TASC), et al. Association scan of 14,500 nonsynonymous SNPs in four diseases identifies autoimmunity variants. Nat Genet. 2007 Nov; 39(11):1329-37. * Member of the consortium and T2D analysis subgroup. PMID: 17952073. , 2007 .

[38]  E. L. Persons,et al.  Ankylosing Spondylitis , 1955, GP.

[39]  N. McElvaney,et al.  Secretory Leucocyte Protease Inhibitor Inhibits Interferon-γ-induced Cathepsin S Expression* , 2007, Journal of Biological Chemistry.

[40]  A. McMichael,et al.  Cell-surface expression and immune receptor recognition of HLA-B27 homodimers. , 2002, Arthritis and rheumatism.

[41]  Dominique Baeten,et al.  Tumor necrosis factor alpha blockade treatment down-modulates the increased systemic and local expression of Toll-like receptor 2 and Toll-like receptor 4 in spondylarthropathy. , 2005, Arthritis and rheumatism.

[42]  J. Taurog.  The Role of HLA-B27 in Spondyloarthritis , 2010, The Journal of Rheumatology.

[43]  E. Schwartz,et al.  Expression of the angiogenic factor thymidine phosphorylase in THP-1 monocytes: induction by autocrine tumor necrosis factor-alpha and inhibition by aspirin. , 2003, Molecular pharmacology.

[44]  Simon C. Potter,et al.  Association scan of 14,500 nonsynonymous SNPs in four diseases identifies autoimmunity variants , 2007, Nature Genetics.

[45]  K. Pahan,et al.  Up-regulation of Microglial CD11b Expression by Nitric Oxide* , 2006, Journal of Biological Chemistry.

[46]  M. Liang,et al.  The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. , 1988, Arthritis and rheumatism.

[47]  P. Zhu,et al.  Identification of disease-associated proteins by proteomic approach in ankylosing spondylitis. , 2007, Biochemical and biophysical research communications.