Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase: A potential mechanism for breaching immunologic tolerance in rheumatoid arthritis

Anne Marie Quirke, Elena Birgitta Lugli, Natalia Wegner, Bart C. Hamilton, Peter Charles, Muslima Chowdhury, A. Jimmy Ytterberg, Roman A. Zubarev, Jan Potempa, Shauna Culshaw, Yonghua Guo, Benjamin A. Fisher, Geoffrey Milton Thiele, Ted R Mikuls, Patrick J W Venables

Research output: Contribution to journalArticle

102 Citations (Scopus)

Abstract

Background Rheumatoid arthritis (RA) is characterised by autoimmunity to citrullinated proteins, and there is increasing epidemiologic evidence linking Porphyromonas gingivalis to RA. P gingivalis is apparently unique among periodontal pathogens in possessing a citrullinating enzyme, peptidylarginine deiminase (PPAD) with the potential to generate antigens driving the autoimmune response. Objectives To examine the immune response to PPAD in patients with RA, individuals with periodontitis (PD) and controls (without arthritis), confirm PPAD autocitrullination and identify the modified arginine residues. Methods PPAD and an inactivated mutant (C351A) were cloned and expressed and autocitrullination of both examined by immunoblotting and mass spectrometry. ELISAs using PPAD, C351A and another P gingivalis protein arginine gingipain (RgpB) were developed and antibody reactivities examined in patients with RA (n=80), individuals with PD (n=44) and controls (n=82). Results Recombinant PPAD was a potent citrullinating enzyme. Antibodies to PPAD, but not to Rgp, were elevated in the RA sera (median 122 U/ml) compared with controls (median 70 U/ml; p<0.05) and PD (median 60 U/ml; p<0.01). Specificity of the anti-peptidyl citrullinated PPAD response was confirmed by the reaction of RA sera with multiple epitopes tested with synthetic citrullinated peptides spanning the PPAD molecule. The elevated antibody response to PPAD was abolished in RA sera if the C351A mutant was used on ELISA. Conclusions The peptidyl citrulline-specific immune response to PPAD supports the hypothesis that, as a bacterial protein, it might break tolerance in RA, and could be a target for therapy.

Original languageEnglish (US)
Pages (from-to)263-269
Number of pages7
JournalAnnals of the rheumatic diseases
Volume73
Issue number1
DOIs
StatePublished - Jan 1 2014

Fingerprint

Porphyromonas gingivalis
Rheumatoid Arthritis
Periodontitis
Autoimmunity
Arginine
Antibodies
protein-arginine deiminase
Serum
Enzyme-Linked Immunosorbent Assay
Citrulline
Bacterial Proteins
Pathogens
Enzymes
Immunoblotting
Arthritis
Antibody Formation
Mass spectrometry
Epitopes
Mass Spectrometry
Proteins

ASJC Scopus subject areas

  • Immunology and Allergy
  • Rheumatology
  • Immunology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase : A potential mechanism for breaching immunologic tolerance in rheumatoid arthritis. / Quirke, Anne Marie; Lugli, Elena Birgitta; Wegner, Natalia; Hamilton, Bart C.; Charles, Peter; Chowdhury, Muslima; Ytterberg, A. Jimmy; Zubarev, Roman A.; Potempa, Jan; Culshaw, Shauna; Guo, Yonghua; Fisher, Benjamin A.; Thiele, Geoffrey Milton; Mikuls, Ted R; Venables, Patrick J W.

In: Annals of the rheumatic diseases, Vol. 73, No. 1, 01.01.2014, p. 263-269.

Research output: Contribution to journalArticle

Quirke, AM, Lugli, EB, Wegner, N, Hamilton, BC, Charles, P, Chowdhury, M, Ytterberg, AJ, Zubarev, RA, Potempa, J, Culshaw, S, Guo, Y, Fisher, BA, Thiele, GM, Mikuls, TR & Venables, PJW 2014, 'Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase: A potential mechanism for breaching immunologic tolerance in rheumatoid arthritis', Annals of the rheumatic diseases, vol. 73, no. 1, pp. 263-269. https://doi.org/10.1136/annrheumdis-2012-202726
Quirke, Anne Marie ; Lugli, Elena Birgitta ; Wegner, Natalia ; Hamilton, Bart C. ; Charles, Peter ; Chowdhury, Muslima ; Ytterberg, A. Jimmy ; Zubarev, Roman A. ; Potempa, Jan ; Culshaw, Shauna ; Guo, Yonghua ; Fisher, Benjamin A. ; Thiele, Geoffrey Milton ; Mikuls, Ted R ; Venables, Patrick J W. / Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase : A potential mechanism for breaching immunologic tolerance in rheumatoid arthritis. In: Annals of the rheumatic diseases. 2014 ; Vol. 73, No. 1. pp. 263-269.
@article{6d056a7df265488383a0a51811191f6c,
title = "Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase: A potential mechanism for breaching immunologic tolerance in rheumatoid arthritis",
abstract = "Background Rheumatoid arthritis (RA) is characterised by autoimmunity to citrullinated proteins, and there is increasing epidemiologic evidence linking Porphyromonas gingivalis to RA. P gingivalis is apparently unique among periodontal pathogens in possessing a citrullinating enzyme, peptidylarginine deiminase (PPAD) with the potential to generate antigens driving the autoimmune response. Objectives To examine the immune response to PPAD in patients with RA, individuals with periodontitis (PD) and controls (without arthritis), confirm PPAD autocitrullination and identify the modified arginine residues. Methods PPAD and an inactivated mutant (C351A) were cloned and expressed and autocitrullination of both examined by immunoblotting and mass spectrometry. ELISAs using PPAD, C351A and another P gingivalis protein arginine gingipain (RgpB) were developed and antibody reactivities examined in patients with RA (n=80), individuals with PD (n=44) and controls (n=82). Results Recombinant PPAD was a potent citrullinating enzyme. Antibodies to PPAD, but not to Rgp, were elevated in the RA sera (median 122 U/ml) compared with controls (median 70 U/ml; p<0.05) and PD (median 60 U/ml; p<0.01). Specificity of the anti-peptidyl citrullinated PPAD response was confirmed by the reaction of RA sera with multiple epitopes tested with synthetic citrullinated peptides spanning the PPAD molecule. The elevated antibody response to PPAD was abolished in RA sera if the C351A mutant was used on ELISA. Conclusions The peptidyl citrulline-specific immune response to PPAD supports the hypothesis that, as a bacterial protein, it might break tolerance in RA, and could be a target for therapy.",
author = "Quirke, {Anne Marie} and Lugli, {Elena Birgitta} and Natalia Wegner and Hamilton, {Bart C.} and Peter Charles and Muslima Chowdhury and Ytterberg, {A. Jimmy} and Zubarev, {Roman A.} and Jan Potempa and Shauna Culshaw and Yonghua Guo and Fisher, {Benjamin A.} and Thiele, {Geoffrey Milton} and Mikuls, {Ted R} and Venables, {Patrick J W}",
year = "2014",
month = "1",
day = "1",
doi = "10.1136/annrheumdis-2012-202726",
language = "English (US)",
volume = "73",
pages = "263--269",
journal = "Annals of the Rheumatic Diseases",
issn = "0003-4967",
publisher = "BMJ Publishing Group",
number = "1",

}

TY - JOUR

T1 - Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase

T2 - A potential mechanism for breaching immunologic tolerance in rheumatoid arthritis

AU - Quirke, Anne Marie

AU - Lugli, Elena Birgitta

AU - Wegner, Natalia

AU - Hamilton, Bart C.

AU - Charles, Peter

AU - Chowdhury, Muslima

AU - Ytterberg, A. Jimmy

AU - Zubarev, Roman A.

AU - Potempa, Jan

AU - Culshaw, Shauna

AU - Guo, Yonghua

AU - Fisher, Benjamin A.

AU - Thiele, Geoffrey Milton

AU - Mikuls, Ted R

AU - Venables, Patrick J W

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Background Rheumatoid arthritis (RA) is characterised by autoimmunity to citrullinated proteins, and there is increasing epidemiologic evidence linking Porphyromonas gingivalis to RA. P gingivalis is apparently unique among periodontal pathogens in possessing a citrullinating enzyme, peptidylarginine deiminase (PPAD) with the potential to generate antigens driving the autoimmune response. Objectives To examine the immune response to PPAD in patients with RA, individuals with periodontitis (PD) and controls (without arthritis), confirm PPAD autocitrullination and identify the modified arginine residues. Methods PPAD and an inactivated mutant (C351A) were cloned and expressed and autocitrullination of both examined by immunoblotting and mass spectrometry. ELISAs using PPAD, C351A and another P gingivalis protein arginine gingipain (RgpB) were developed and antibody reactivities examined in patients with RA (n=80), individuals with PD (n=44) and controls (n=82). Results Recombinant PPAD was a potent citrullinating enzyme. Antibodies to PPAD, but not to Rgp, were elevated in the RA sera (median 122 U/ml) compared with controls (median 70 U/ml; p<0.05) and PD (median 60 U/ml; p<0.01). Specificity of the anti-peptidyl citrullinated PPAD response was confirmed by the reaction of RA sera with multiple epitopes tested with synthetic citrullinated peptides spanning the PPAD molecule. The elevated antibody response to PPAD was abolished in RA sera if the C351A mutant was used on ELISA. Conclusions The peptidyl citrulline-specific immune response to PPAD supports the hypothesis that, as a bacterial protein, it might break tolerance in RA, and could be a target for therapy.

AB - Background Rheumatoid arthritis (RA) is characterised by autoimmunity to citrullinated proteins, and there is increasing epidemiologic evidence linking Porphyromonas gingivalis to RA. P gingivalis is apparently unique among periodontal pathogens in possessing a citrullinating enzyme, peptidylarginine deiminase (PPAD) with the potential to generate antigens driving the autoimmune response. Objectives To examine the immune response to PPAD in patients with RA, individuals with periodontitis (PD) and controls (without arthritis), confirm PPAD autocitrullination and identify the modified arginine residues. Methods PPAD and an inactivated mutant (C351A) were cloned and expressed and autocitrullination of both examined by immunoblotting and mass spectrometry. ELISAs using PPAD, C351A and another P gingivalis protein arginine gingipain (RgpB) were developed and antibody reactivities examined in patients with RA (n=80), individuals with PD (n=44) and controls (n=82). Results Recombinant PPAD was a potent citrullinating enzyme. Antibodies to PPAD, but not to Rgp, were elevated in the RA sera (median 122 U/ml) compared with controls (median 70 U/ml; p<0.05) and PD (median 60 U/ml; p<0.01). Specificity of the anti-peptidyl citrullinated PPAD response was confirmed by the reaction of RA sera with multiple epitopes tested with synthetic citrullinated peptides spanning the PPAD molecule. The elevated antibody response to PPAD was abolished in RA sera if the C351A mutant was used on ELISA. Conclusions The peptidyl citrulline-specific immune response to PPAD supports the hypothesis that, as a bacterial protein, it might break tolerance in RA, and could be a target for therapy.

UR - http://www.scopus.com/inward/record.url?scp=84889663283&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84889663283&partnerID=8YFLogxK

U2 - 10.1136/annrheumdis-2012-202726

DO - 10.1136/annrheumdis-2012-202726

M3 - Article

C2 - 23463691

AN - SCOPUS:84889663283

VL - 73

SP - 263

EP - 269

JO - Annals of the Rheumatic Diseases

JF - Annals of the Rheumatic Diseases

SN - 0003-4967

IS - 1

ER -