HLA–B27–restricted CD8+ T cell response to cartilage-derived self peptides in ankylosing spondylitis
Pamir Atagunduz
Charité University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany
Search for more papers by this authorHeiner Appel
Charité University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany
Search for more papers by this authorWolfgang Kuon
Charité University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany
Search for more papers by this authorPeihua Wu
Charité University Medicine Berlin, Campus Benjamin Franklin, and German Rheumatology Research Center, Berlin, Germany
Search for more papers by this authorAndreas Thiel
German Rheumatology Research Center, Berlin, Germany
Search for more papers by this authorCorresponding Author
Joachim Sieper
Charité University Medicine Berlin, Campus Benjamin Franklin, and German Rheumatology Research Center, Berlin, Germany
Medical Department I, Rheumatology, Benjamin Franklin University Hospital, Hindenburgdamm 30, 12200 Berlin, GermanySearch for more papers by this authorPamir Atagunduz
Charité University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany
Search for more papers by this authorHeiner Appel
Charité University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany
Search for more papers by this authorWolfgang Kuon
Charité University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany
Search for more papers by this authorPeihua Wu
Charité University Medicine Berlin, Campus Benjamin Franklin, and German Rheumatology Research Center, Berlin, Germany
Search for more papers by this authorAndreas Thiel
German Rheumatology Research Center, Berlin, Germany
Search for more papers by this authorCorresponding Author
Joachim Sieper
Charité University Medicine Berlin, Campus Benjamin Franklin, and German Rheumatology Research Center, Berlin, Germany
Medical Department I, Rheumatology, Benjamin Franklin University Hospital, Hindenburgdamm 30, 12200 Berlin, GermanySearch for more papers by this authorAbstract
Objective
Several hypotheses have been proposed to explain the strong association between HLA–B27 and ankylosing spondylitis (AS). Among these, the arthritogenic peptide theory proposes that certain B27 subtype alleles bind specific arthritogenic peptide(s) due to their unique amino acid anchor residues. Cartilage antigens have been discussed as candidate targets for the immune response in AS. The recognition of HLA–B27–peptide complexes by self-reactive CD8+ T cells might contribute to joint-specific tissue damage. Therefore, we investigated the presence of autoreactive CD8+ T cells specific for cartilage-derived peptides in patients with AS.
Methods
An HLA–B27–binding prediction program and a proteasome-cutting prediction program for the human 20S proteasome were used to screen 18 human cartilage proteins for potentially immunogenic nonamer peptides. The peptides identified were used to stimulate peripheral blood mononuclear cells from 20 HLA–B27–positive patients with AS and synovial fluid (SF) mononuclear cells from 7 HLA–B27–positive patients with AS. Activation of T cells was measured by antigen-specific intracellular cytokine staining and quantified by flow cytometry.
Results
From the screening analysis, we identified 121 nonamer peptides. Of these, 1 peptide derived from type II collagen and 1 from type VI collagen were stimulatory for peripheral blood CD8+ T cells in only 1 of 20 patients. However, in 4 of 7 SF samples the same type VI collagen–derived nonamer peptide stimulated SF CD8+ T cells, but none of the other peptides was stimulatory. This CD8+ T cell response could be blocked by an anti–HLA–B27 antibody, confirming an HLA–B27–restricted immune response.
Conclusion
Our findings suggest that cartilage-directed cellular autoimmunity might play an important role in joint-specific tissue damage in patients with AS. Future research is necessary to determine whether the identified peptide is of pathogenetic relevance.
REFERENCES
- 1 Braun J, Bollow M, Remlinger G, Eggens U, Rudwaleit M, Distler A, et al. Prevalence of spondylarthropathies in HLA–B27 positive and negative blood donors. Arthritis Rheum 1998; 41: 58–67.
- 2 Marker-Hermann E, Hoehler T. Pathogenesis of human leukocyte antigen B27-positive arthritis: information from clinical materials. Rheum Dis Clin North Am 1998; 24: 865–81.
- 3 Khan MA. HLA-B27 polymorphism and association with disease. J Rheumatol 2000; 27: 1110–4.
- 4 Kuon W, Sieper J. Identification of HLA-B27-restricted peptides in reactive arthritis and other spondyloarthropathies: computer algorithms and fluorescent activated cell sorting analysis as tools for hunting of HLA-B27-restricted chlamydial and autologous crossreactive peptides involved in reactive arthritis and ankylosing spondylitis. Rheum Dis Clin North Am 2003; 29: 595–611.
- 5 D'Amato M, Fiorillo MT, Carcassi C, Mathieu A, Zuccarelli A, Bitti PP, et al. Relevance of residue 116 of HLA-B27 in determining susceptibility to ankylosing spondylitis. Eur J Immunol 1995; 25: 3199–201.
- 6 Lopez-Larrea C, Sujirachato K, Mehra NK, Chiewsilp P, Isarangkura D, Kanga U, et al. HLA-B27 subtypes in Asian patients with ankylosing spondylitis: evidence for new associations. Tissue Antigens 1995; 45: 169–76.
- 7 Nasution AR, Mardjuadi A, Kunmartini S, Suryadhana NG, Setyohadi B, Sudarsono D, et al. HLA-B27 subtypes positively and negatively associated with spondyloarthropathy. J Rheumatol 1997; 24: 1111–4.
- 8 Ren EC, Koh WH, Sim D, Boey ML, Wee GB, Chan SH. Possible protective role of HLA-B*2706 for ankylosing spondylitis. Tissue Antigens 1997; 49: 67–9.
- 9 Kuon W, Holzhutter HG, Appel H, Grolms M, Kollnberger S, Traeder A, et al. Identification of HLA-B27-restricted peptides from the Chlamydia trachomatis proteome with possible relevance to HLA-B27-associated diseases. J Immunol 2001; 167: 4738–46.
- 10 Huang F, Hermann E, Wang J, Cheng XK, Tsai WC, Wen J, et al. A patient-derived cytotoxic T-lymphocyte clone and two peptide-dependent monoclonal antibodies recognize HLA-B27-peptide complexes with low stringency for peptide sequences. Infect Immun 1996; 64: 120–7.
- 11 Ugrinovic S, Mertz A, Wu P, Braun J, Sieper J. A single nonamer from the Yersinia 60-kDa heat shock protein is the target of HLA-B27-restricted CTL response in Yersinia-induced reactive arthritis. J Immunol 1997; 159: 5715–23.
- 12 May E, Dulphy N, Frauendorf E, Duchmann R, Bowness P, Lopez de Castro JA, et al. Conserved TCR β chain usage in reactive arthritis: evidence for selection by a putative HLA-B27-associated autoantigen. Tissue Antigens 2002; 60: 299–308.
- 13 Hermann E, Yu DT, Meyer zum Buschenfelde KH, Fleischer B. HLA-B27-restricted CD8 T cells derived from synovial fluids of patients with reactive arthritis and ankylosing spondylitis. Lancet 1993; 342: 646–50.
- 14 Hermann E, Sucke B, Droste U, Meyer zum Buschenfelde KH. Klebsiella pneumoniae–reactive T cells in blood and synovial fluid of patients with ankylosing spondylitis: comparison with HLA–B27+ healthy control subjects in a limiting dilution study and determination of the specificity of synovial fluid T cell clones. Arthritis Rheum 1995; 38: 1277–82.
- 15 Frauendorf E, von Goessel H, May E, Marker-Hermann E. HLA-B27-restricted T cells from patients with ankylosing spondylitis recognize peptides from B*2705 that are similar to bacteria-derived peptides. Clin Exp Immunol 2003; 134: 351–9.
- 16 McGonagle D, Gibbon W, Emery P. Classification of inflammatory arthritis by enthesitis. Lancet 1998; 352: 1137–40.
- 17 Bollow M, Fischer T, Reisshauer H, Backhaus M, Sieper J, Hamm B, et al. Quantitative analysis of sacroiliac biopsies in spondyloarthropathies: T cells and macrophages predominate in early and active sacroiliitis: cellularity correlates with the degree of enhancement detected by magnetic resonance imaging. Ann Rheum Dis 2000; 59: 135–40.
- 18 Braun J, Bollow M, Eggens U, Konig H, Distler A, Sieper J. Use of dynamic magnetic resonance imaging with fast imaging in the detection of early and advanced sacroiliitis in spondylarthropathy patients. Arthritis Rheum 1994; 37: 1039–45.
- 19 McGonagle D, Conaghan PG, O'Connor P, Gibbon W, Green M, Wakefield R, et al. The relationship between synovitis and bone changes in early untreated rheumatoid arthritis: a controlled magnetic resonance imaging study. Arthritis Rheum 1999; 42: 1706–11.
- 20 Braun J, Bollow M, Neure L, Seipelt E, Seyrekbasan F, Herbst H, et al. Use of immunohistologic and in situ hybridization techniques in the examination of sacroiliac joint biopsy specimens from patients with ankylosing spondylitis. Arthritis Rheum 1995; 38: 499–505.
- 21 Francois RJ, Gardner DL, Degrave EJ, Bywaters EG. Histopathologic evidence that sacroiliitis in ankylosing spondylitis is not merely enthesitis: systematic study of specimens from patients and control subjects. Arthritis Rheum 2000; 43: 2011–24.
- 22 Poole AR. The histopathology of ankylosing spondylitis: are there unifying hypotheses? Am J Med Sci 1998; 316: 228–33.
- 23 Maksymowych WP. Ankylosing spondylitis: at the interface of bone and cartilage. J Rheumatol 2000; 27: 2295–301.
- 24 Braun J, Khan MA, Sieper J. Enthesitis and ankylosis in spondyloarthropathy: what is the target of the immune response? Ann Rheum Dis 2000; 59: 985–94.
- 25 Zhang Y, Guerassimov A, Leroux JY, Cartman A, Webber C, Lalic R, et al. Arthritis induced by proteoglycan aggrecan G1 domain in BALB/c mice: evidence for T cell involvement and the immunosuppressive influence of keratan sulfate on recognition of T and B cell epitopes. J Clin Invest 1998; 101: 1678–86.
- 26 Mikecz K, Glant TT, Baron M, Poole AR. Isolation of proteoglycan-specific T lymphocytes from patients with ankylosing spondylitis. Cell Immunol 1988; 112: 55–63.
- 27 Guerassimov A, Zhang Y, Cartman A, Rosenberg LC, Esdaile J, Fitzcharles MA, et al. Immune responses to cartilage link protein and the G1 domain of proteoglycan aggrecan in patients with osteoarthritis. Arthritis Rheum 1999; 42: 527–33.
- 28 Zou J, Zhang Y, Thiel A, Rudwaleit M, Shi SL, Radbruch A, et al. Predominant cellular immune response to the cartilage autoantigenic G1 aggrecan in ankylosing spondylitis and rheumatoid arthritis. Rheumatology (Oxford) 2003; 42: 846–55.
- 29 Zou J, Appel H, Rudwaleit M, Thiel A, Sieper J. Analysis of the CD8+ T cell response to the G1 domain of aggregan in ankylosing spondylitis. Ann Rheum Dis 2004 [Epub ahead of print].
- 30 Gao XM, Wordsworth P, McMichael A. Collagen-specific cytotoxic T lymphocyte responses in patients with ankylosing spondylitis and reactive arthritis. Eur J Immunol 1994; 24: 1665–70.
- 31 Van der Linden S, Valkenburg HA, Cats A. Evaluation of diagnostic criteria for ankylosing spondylitis: a proposal for modification of the New York criteria. Arthritis Rheum 1984; 27: 361–8.
- 32 Rammensee H, Bachmann J, Emmerich NP, Bachor OA, Stevanovic S. SYFPEITHI: database for MHC ligands and peptide motifs. Immunogenetics 1999; 50: 213–9.
- 33 Holzhutter HG, Frommel C, Kloetzel PM. A theoretical approach towards the identification of cleavage: determining amino acid motifs of the 20S proteasome. J Mol Biol 1999; 286: 1251–65.
- 34 Holzhutter HG, Kloetzel PM. A kinetic model of vertebrate 20S proteasome accounting for the generation of major proteolytic fragments from oligomeric peptide substrates. Biophys J 2000; 79: 1196–205.
- 35 Jung G, Beck-Sickinger AG. Multiple peptide synthesis methods and their applications. Angew Chem Int Ed Engl 1992; 31: 367–83.
- 36 Thiel A, Radbruch A. Antigen-specific cytometry. Arthritis Res 1999; 1: 25–9.
- 37 Thiel A, Wu P, Lauster R, Braun J, Radbruch A, Sieper J. Analysis of the antigen-specific T cell response in reactive arthritis by flow cytometry. Arthritis Rheum 2000; 43: 2834–42.
- 38 Thiel A, Scheffold A, Radbruch A. Antigen-specific flow cytometry: new tools arrived. Clin Immunol 2004; 111: 155–61.
- 39 Waldrop SL, Pitcher CJ, Peterson DM, Maino VC, Picker LJ. Determination of antigen-specific memory/effector CD4+ T cell frequencies by flow cytometry: evidence for a novel, antigen-specific homeostatic mechanism in HIV-associated immunodeficiency. J Clin Invest 1997; 99: 1739–50.
- 40 Brooks JM, Murray RJ, Thomas WA, Kurilla MG, Rickinson AB. Different HLA-B27 subtypes present the same immunodominant Epstein-Barr virus peptide. J Exp Med 1993; 178: 879–87.
- 41 Tan TL, Geluk A, Toebes M, Ottenhoff TH, Drijfhout JW. A novel, highly efficient peptide-HLA class I binding assay using unfolded heavy chain molecules: identification of HIV-1 derived peptides that bind to HLA-A*0201 and HLA-A*0301. J Immunol Methods 1997; 205: 201–9.
- 42 Rotzschke O, Falk K, Stevanovic S, Gnau V, Jung G, Rammensee HG. Dominant aromatic/aliphatic C-terminal anchor in HLA-B*2702 and B*2705 peptide motifs. Immunogenetics 1994; 39: 74–7.
- 43 Fiorillo MT, Maragno M, Butler R, Dupuis ML, Sorrentino R. CD8+ T-cell autoreactivity to an HLA-B27-restricted self-epitope correlates with ankylosing spondylitis. J Clin Invest 2000; 106: 47–53.
- 44 Poole CA, Flint MH, Beaumont BW. Chondrons in cartilage: ultrastructural analysis of the pericellular microenvironment in adult human articular cartilage. J Orthop Res 1987; 5: 191–7.
- 45 Poole CA, Ayad S, Gilbert RT. Chondrons from articular cartilage: immunohistochemical evaluation of type VI collagen organisation in isolated chondrons by light, confocal and electron microscopy. J Cell Sci 1992; 103: 1101–10.
- 46 Townsend A, Ohlen C, Bastin J, Ljunggren HG, Foster L, Karre K. Association of class I major histocompatibility heavy and light chains induced by viral peptides. Nature 1989; 340: 443–8.
- 47 Dedier S, Reinelt S, Reitinger T, Folkers G, Rognan D. Thermodynamic stability of HLA-B*2705 peptide complexes: effect of peptide and major histocompatibility complex protein mutations. J Biol Chem 2000; 275: 27055–61.
- 48 Lopez de Castro JA, Alvarez I, Marcilla M, Paradela A, Ramos M, Sesma L, et al. HLA-B27: a registry of constitutive peptide ligands. Tissue Antigens 2004; 63: 424–45.
- 49 Andersen ML, Ruhwald M, Nissen MH, Buus S, Claesson MH. Self-peptides with intermediate capacity to bind and stabilize MHC class I molecules may be immunogenic. Scand J Immunol 2003; 57: 21–7.
- 50 Taurog JD, Richardson JA, Croft JT, Simmons WA, Zhou M, Fernandez-Sueiro JL, et al. The germfree state prevents development of gut and joint inflammatory disease in HLA-B27 transgenic rats. J Exp Med 1994; 180: 2359–64.
- 51 Sieper J, Braun J. Pathogenesis of spondylarthropathies: persistent bacterial antigen, autoimmunity, or both? Arthritis Rheum 1995; 38: 1547–54.
- 52 Fan L, Busser BW, Lifsted TQ, Oukka M, Lo D, Laufer TM. Antigen presentation by keratinocytes directs autoimmune skin disease. Proc Natl Acad Sci U S A 2003; 100: 3386–91.
- 53 Savinov AY, Wong FS, Stonebraker AC, Chervonsky AV. Presentation of antigen by endothelial cells and chemoattraction are required for homing of insulin-specific CD8+ T cells. J Exp Med 2003; 197: 643–56.
- 54 Mack CL, Vanderlugt-Castaneda CL, Neville KL, Miller SD. Microglia are activated to become competent antigen presenting and effector cells in the inflammatory environment of the Theiler's virus model of multiple sclerosis. J Neuroimmunol 2003; 144: 68–79.
- 55 Cohen ES, Bodmer HC. Cytotoxic T lymphocytes recognize and lyse chondrocytes under inflammatory, but not non-inflammatory conditions. Immunology 2003; 109: 8–14.
