Identification of an immunodominant peptide from citrullinated tenascin-C as a major target for autoantibodies in rheumatoid arthritis

Objectives We investigated whether citrullinated tenascin-C (cTNC), an extracellular matrix protein expressed at high levels in the joints of patients with rheumatoid arthritis (RA), is a target for the autoantibodies in RA. Methods Citrullinated sites were mapped by mass spectrometry in the fibrinogen-like globe (FBG) domain of tenascin-C treated with peptidylarginine deiminases (PAD) 2 and 4. Antibodies to cyclic peptides containing citrullinated sites were screened in sera from patients with RA by ELISA. Potential cross-reactivity with well-established anticitrullinated protein antibody (ACPA) epitopes was tested by inhibition assays. The autoantibody response to one immunodominant cTNC peptide was then analysed in 101 pre-RA sera (median 7 years before onset) and two large independent RA cohorts. Results Nine arginine residues within FBG were citrullinated by PAD2 and PAD4. Two immunodominant peptides cTNC1 (VFLRRKNG-cit-ENFYQNW) and cTNC5 (EHSIQFAEMKL-cit-PSNF-cit-NLEG-cit-cit-KR) were identified. Antibodies to both showed limited cross-reactivity with ACPA epitopes from α-enolase, vimentin and fibrinogen, and no reactivity with citrullinated fibrinogen peptides sharing sequence homology with FBG. cTNC5 antibodies were detected in 18% of pre-RA sera, and in 47% of 1985 Swedish patients with RA and 51% of 287 North American patients with RA. The specificity was 98% compared with 160 healthy controls and 330 patients with osteoarthritis. Conclusions There are multiple citrullination sites in the FBG domain of tenascin-C. Among these, one epitope is recognised by autoantibodies that are detected years before disease onset, and which may serve as a useful biomarker to identify ACPA-positive patients with high sensitivity and specificity in established disease.

[1]  W. Yue,et al.  Crystal structure of Porphyromonas gingivalis peptidylarginine deiminase: implications for autoimmunity in rheumatoid arthritis , 2015, Annals of the rheumatic diseases.

[2]  L. Klareskog,et al.  Autoimmunity: Lungs and citrullination , 2015, Nature Reviews Rheumatology.

[3]  R. Fischer,et al.  Gel-aided sample preparation (GASP)—A simplified method for gel-assisted proteomic sample generation from protein extracts and intact cells , 2015, Proteomics.

[4]  R. Fischer,et al.  Expression of citrulline and homocitrulline residues in the lungs of non-smokers and smokers: implications for autoimmunity in rheumatoid arthritis , 2015, Arthritis Research & Therapy.

[5]  W. Robinson,et al.  Periodontitis and Porphyromonas gingivalis in Patients With Rheumatoid Arthritis , 2014, Arthritis & rheumatology.

[6]  G. D. de Souza,et al.  Assessing the citrullinome in rheumatoid arthritis synovial fluid with and without enrichment of citrullinated peptides. , 2014, Journal of proteome research.

[7]  J. Drijfhout,et al.  The human peptidylarginine deiminases type 2 and type 4 have distinct substrate specificities. , 2014, Biochimica et biophysica acta.

[8]  P. Vineis,et al.  O19. Smoking Affects Immune Response to Citrullinated Autoantigens Before Clinical Onset of Rheumatoid Arthritis in a Southern European Cohort , 2014 .

[9]  W. Robinson,et al.  Identification of anticitrullinated protein antibody reactivities in a subset of anti-CCP-negative rheumatoid arthritis: association with cigarette smoking and HLA-DRB1 ‘shared epitope’ alleles , 2013, Annals of the rheumatic diseases.

[10]  M. Chiquet,et al.  Cleavage of extracellular matrix in periodontitis: gingipains differentially affect cell adhesion activities of fibronectin and tenascin-C. , 2013, Biochimica et biophysica acta.

[11]  G. Herrero-Beaumont,et al.  Citrullination enhances the pro-inflammatory response to fibrin in rheumatoid arthritis synovial fibroblasts , 2012, Annals of the rheumatic diseases.

[12]  P. Taylor,et al.  Raised circulating tenascin-C in rheumatoid arthritis , 2012, Arthritis Research & Therapy.

[13]  K. Midwood,et al.  Endogenous Control of Immunity against Infection: Tenascin-C Regulates TLR4-Mediated Inflammation via MicroRNA-155 , 2012, Cell reports.

[14]  K. Midwood,et al.  Endogenous activation of adaptive immunity: tenascin-C drives interleukin-17 synthesis in murine arthritic joint disease. , 2012, Arthritis and rheumatism.

[15]  L. Alfredsson,et al.  Genetic and environmental determinants for disease risk in subsets of rheumatoid arthritis defined by the anticitrullinated protein/peptide antibody fine specificity profile , 2012, Annals of the rheumatic diseases.

[16]  R. Toes,et al.  Induction of osteoclastogenesis and bone loss by human autoantibodies against citrullinated vimentin. , 2012, The Journal of clinical investigation.

[17]  Peter J Sterk,et al.  Extracellular matrix composition in COPD , 2012, European Respiratory Journal.

[18]  J. Drijfhout,et al.  Anti-citrullinated fibronectin antibodies in rheumatoid arthritis are associated with human leukocyte antigen-DRB1 shared epitope alleles , 2012, Arthritis Research & Therapy.

[19]  R. Kaarteenaho,et al.  Tenascin-C and alpha-smooth muscle actin positive cells are increased in the large airways in patients with COPD , 2011, Respiratory research.

[20]  Y. Matsui,et al.  α9β1 Integrin-Mediated Signaling Serves as an Intrinsic Regulator of Pathogenic Th17 Cell Generation , 2011, The Journal of Immunology.

[21]  D. Symmons,et al.  Antibodies to citrullinated α-enolase peptide 1 and clinical and radiological outcomes in rheumatoid arthritis , 2011, Annals of the rheumatic diseases.

[22]  R. Schasfoort,et al.  Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance , 2010, Arthritis research & therapy.

[23]  P. Nigrovic,et al.  Protection against anti-citrullinated protein antibody-positive rheumatoid arthritis is predominantly associated with HLA-DRB1*1301: A meta-analysis of HLA-DRB1 associations with anti-citrullinated protein antibody-positive and anti-citrullinated protein antibody-negative rheumatoid arthritis in fou , 2010 .

[24]  R. Wait,et al.  Peptidylarginine deiminase from Porphyromonas gingivalis citrullinates human fibrinogen and α-enolase: implications for autoimmunity in rheumatoid arthritis. , 2010, Arthritis and rheumatism.

[25]  Lars Alfredsson,et al.  Protection against anti-citrullinated protein antibody-positive rheumatoid arthritis is predominantly associated with HLA-DRB1*1301: a meta-analysis of HLA-DRB1 associations with anti-citrullinated protein antibody-positive and anti-citrullinated protein antibody-negative rheumatoid arthritis in fou , 2010, Arthritis and rheumatism.

[26]  I. Udalova,et al.  Transcriptional Regulation of the Endogenous Danger Signal Tenascin-C: A Novel Autocrine Loop in Inflammation , 2010, The Journal of Immunology.

[27]  M. Feldmann,et al.  Autoimmunity to specific citrullinated proteins gives the first clues to the etiology of rheumatoid arthritis , 2010, Immunological reviews.

[28]  Lars Alfredsson,et al.  Specific interaction between genotype, smoking and autoimmunity to citrullinated α-enolase in the etiology of rheumatoid arthritis , 2009, Nature Genetics.

[29]  K. Midwood,et al.  J. Cell Commun. Signal. (2009) 3:287–310 DOI 10.1007/s12079-009-0075-1 RESEARCH ARTICLE The role of tenascin-C in tissue injury and tumorigenesis , 2009 .

[30]  C. Benoist,et al.  A broad screen for targets of immune complexes decorating arthritic joints highlights deposition of nucleosomes in rheumatoid arthritis , 2009, Proceedings of the National Academy of Sciences.

[31]  M. Kashiwagi,et al.  Tenascin-C is an endogenous activator of Toll-like receptor 4 that is essential for maintaining inflammation in arthritic joint disease , 2009, Nature Medicine.

[32]  R. Muschel,et al.  Novel MMP-9 Substrates in Cancer Cells Revealed by a Label-free Quantitative Proteomics Approach*S , 2008, Molecular & Cellular Proteomics.

[33]  R. Wait,et al.  Antibodies to citrullinated alpha-enolase peptide 1 are specific for rheumatoid arthritis and cross-react with bacterial enolase. , 2008, Arthritis and rheumatism.

[34]  R. Wait,et al.  Synovial fluid is a site of citrullination of autoantigens in inflammatory arthritis. , 2008, Arthritis and rheumatism.

[35]  J. Lundeberg,et al.  Multiple antibody reactivities to citrullinated antigens in sera from patients with rheumatoid arthritis: association with HLA-DRB1 alleles , 2008, Annals of the rheumatic diseases.

[36]  B. Dijkmans,et al.  Progression of joint damage in early rheumatoid arthritis: association with HLA-DRB1, rheumatoid factor, and anti-citrullinated protein antibodies in relation to different treatment strategies. , 2008, Arthritis and rheumatism.

[37]  M. Sebbag,et al.  Induction of macrophage secretion of tumor necrosis factor alpha through Fcgamma receptor IIa engagement by rheumatoid arthritis-specific autoantibodies to citrullinated proteins complexed with fibrinogen. , 2008, Arthritis and rheumatism.

[38]  P. Gregersen,et al.  Gene-gene and gene-environment interactions involving HLA-DRB1, PTPN22, and smoking in two subsets of rheumatoid arthritis. , 2007, American journal of human genetics.

[39]  A. Sudo,et al.  Expression of large tenascin‐C splice variants in synovial fluid of patients with rheumatoid arthritis , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[40]  W. Robinson,et al.  Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis. , 2006, The Journal of clinical investigation.

[41]  Elizabeth W Karlson,et al.  Replication of putative candidate-gene associations with rheumatoid arthritis in >4,000 samples from North America and Sweden: association of susceptibility with PTPN22, CTLA4, and PADI4. , 2005, American journal of human genetics.

[42]  Wei Chen,et al.  Refining the complex rheumatoid arthritis phenotype based on specificity of the HLA-DRB1 shared epitope for antibodies to citrullinated proteins. , 2005, Arthritis and rheumatism.

[43]  G. Weissmann,et al.  Hypothesis: The Humoral Immune Response to Oral Bacteria Provides a Stimulus for the Development of Rheumatoid Arthritis , 2004, Inflammation.

[44]  P. Tak,et al.  The presence of citrullinated proteins is not specific for rheumatoid synovial tissue. , 2004, Arthritis and rheumatism.

[45]  L. Alfredsson,et al.  A gene-environment interaction between smoking and shared epitope genes in HLA-DR provides a high risk of seropositive rheumatoid arthritis. , 2004, Arthritis and rheumatism.

[46]  E. Lapointe,et al.  Rheumatoid arthritis specific anti-Sa antibodies target citrullinated vimentin , 2004, Arthritis research & therapy.

[47]  B. Dijkmans,et al.  Specific autoantibodies precede the symptoms of rheumatoid arthritis: a study of serial measurements in blood donors. , 2004, Arthritis and rheumatism.

[48]  Göran Hallmans,et al.  Antibodies against cyclic citrullinated peptide and IgA rheumatoid factor predict the development of rheumatoid arthritis. , 2003, Arthritis and rheumatism.

[49]  L. Alfredsson,et al.  Quantification of the influence of cigarette smoking on rheumatoid arthritis: results from a population based case-control study, using incident cases , 2003, Annals of the rheumatic diseases.

[50]  V. Pešáková,et al.  Autoantibodies can be prognostic markers of an erosive disease in early rheumatoid arthritis , 2003, Annals of the rheumatic diseases.

[51]  E. Girbal-Neuhauser,et al.  The Major Synovial Targets of the Rheumatoid Arthritis-Specific Antifilaggrin Autoantibodies Are Deiminated Forms of the α- and β-Chains of Fibrin1 , 2001, The Journal of Immunology.

[52]  M. V. van Leeuwen,et al.  The prognostic value of anti-cyclic citrullinated peptide antibody in patients with recent-onset rheumatoid arthritis. , 2000, Arthritis and rheumatism.

[53]  M. Jolivet,et al.  The epitopes targeted by the rheumatoid arthritis-associated antifilaggrin autoantibodies are posttranslationally generated on various sites of (pro)filaggrin by deimination of arginine residues. , 1999, Journal of immunology.

[54]  A. Silman,et al.  Cigarette smoking increases the risk of rheumatoid arthritis. Results from a nationwide study of disease-discordant twins. , 1996, Arthritis and rheumatism.

[55]  L. Zardi,et al.  Tenascin distribution in articular cartilage from normal subjects and from patients with osteoarthritis and rheumatoid arthritis. , 1994, Arthritis and rheumatism.

[56]  D. Salter,et al.  Tenascin is increased in cartilage and synovium from arthritic knees. , 1993, British journal of rheumatology.

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

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

[59]  J. Sokolove,et al.  Immune complexes containing citrullinated fibrinogen costimulate macrophages via Toll-like receptor 4 and Fcγ receptor. , 2011, Arthritis and rheumatism.

[60]  I. Udalova,et al.  Expression and immune function of tenascin-C. , 2011, Critical reviews in immunology.

[61]  W. Robinson,et al.  Immune Complexes Containing Citrullinated Fibrinogen Costimulate Macrophages via Toll-like Receptor 4 and Fc Receptor , 2010 .

[62]  F. V. D. van den Hoogen,et al.  Citrulline is an essential constituent of antigenic determinants recognized by rheumatoid arthritis-specific autoantibodies. , 1998, The Journal of clinical investigation.