Autoantibodies from Single Circulating Plasmablasts React with Citrullinated Antigens and Porphyromonas gingivalis in Rheumatoid Arthritis

Song Li, Yangsheng Yu, Yinshi Yue, Hongyan Liao, Wanqin Xie, Jessica Thai, Ted R Mikuls, Geoffrey Milton Thiele, Michael J. Duryee, Harlan Sayles, Jeffrey B Payne, Lynell Warren Klassen, James Robert O'Dell, Zhixin Zhang, Kaihong Su

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Objective Anti-citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA). However, the molecular basis for ACPA production is still unclear. The purpose of this study was to determine if circulating plasmablasts from RA patients produce ACPAs and whether Porphyromonas gingivalis facilitates the generation of ACPAs. Methods Using a single-cell antibody cloning approach, we generated 217 and 110 monoclonal recombinant antibodies from circulating plasmablasts from 7 RA patients and 4 healthy controls, respectively. Antibody reactivity with citrullinated antigens was tested by a second-generation anti-cyclic citrullinated peptide (anti-CCP) kit and by enzyme-linked immunosorbent assays (ELISAs) against citrullinated human antigens. Antibody reactivity with P gingivalis was tested by ELISAs against outer membrane antigens (OMAs) and citrullinated enolase from P gingivalis. Results Approximately 19.5% of plasmablast-derived antibodies from anti-CCP-positive RA patients, but none from 1 anti-CCP-negative RA patient or the healthy controls, specifically recognized citrullinated antigens. The immunoglobulin genes encoding these ACPAs were highly mutated, with increased ratios of replacement mutations to silent mutations, suggesting the involvement of active antigen selection in ACPA generation. Interestingly, 63% of the ACPAs cross-reacted with OMAs and/or citrullinated enolase from P gingivalis. The reactivity of ACPAs against citrullinated proteins from P gingivalis was confirmed by immunoblotting and mass spectrometry. Furthermore, some germline-reverted ACPAs retained their reactivity with P gingivalis antigens but completely lost their reactivity with citrullinated human antigens. Conclusion These results suggest that circulating plasmablasts in RA patients produce ACPAs and that this process may be facilitated by anti-P gingivalis immune responses.

Original languageEnglish (US)
Pages (from-to)614-626
Number of pages13
JournalArthritis and Rheumatology
Volume68
Issue number3
DOIs
StatePublished - Mar 1 2016

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Porphyromonas gingivalis
Autoantibodies
Rheumatoid Arthritis
Antigens
Antibodies
Proteins
Phosphopyruvate Hydratase
Enzyme-Linked Immunosorbent Assay
Immunoglobulin Genes
Membranes
Immunoblotting
Antibody Formation
Organism Cloning
Anti-Idiotypic Antibodies
Mass Spectrometry

ASJC Scopus subject areas

  • Immunology and Allergy
  • Rheumatology
  • Immunology

Cite this

Autoantibodies from Single Circulating Plasmablasts React with Citrullinated Antigens and Porphyromonas gingivalis in Rheumatoid Arthritis. / Li, Song; Yu, Yangsheng; Yue, Yinshi; Liao, Hongyan; Xie, Wanqin; Thai, Jessica; Mikuls, Ted R; Thiele, Geoffrey Milton; Duryee, Michael J.; Sayles, Harlan; Payne, Jeffrey B; Klassen, Lynell Warren; O'Dell, James Robert; Zhang, Zhixin; Su, Kaihong.

In: Arthritis and Rheumatology, Vol. 68, No. 3, 01.03.2016, p. 614-626.

Research output: Contribution to journalArticle

Li, Song ; Yu, Yangsheng ; Yue, Yinshi ; Liao, Hongyan ; Xie, Wanqin ; Thai, Jessica ; Mikuls, Ted R ; Thiele, Geoffrey Milton ; Duryee, Michael J. ; Sayles, Harlan ; Payne, Jeffrey B ; Klassen, Lynell Warren ; O'Dell, James Robert ; Zhang, Zhixin ; Su, Kaihong. / Autoantibodies from Single Circulating Plasmablasts React with Citrullinated Antigens and Porphyromonas gingivalis in Rheumatoid Arthritis. In: Arthritis and Rheumatology. 2016 ; Vol. 68, No. 3. pp. 614-626.
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title = "Autoantibodies from Single Circulating Plasmablasts React with Citrullinated Antigens and Porphyromonas gingivalis in Rheumatoid Arthritis",
abstract = "Objective Anti-citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA). However, the molecular basis for ACPA production is still unclear. The purpose of this study was to determine if circulating plasmablasts from RA patients produce ACPAs and whether Porphyromonas gingivalis facilitates the generation of ACPAs. Methods Using a single-cell antibody cloning approach, we generated 217 and 110 monoclonal recombinant antibodies from circulating plasmablasts from 7 RA patients and 4 healthy controls, respectively. Antibody reactivity with citrullinated antigens was tested by a second-generation anti-cyclic citrullinated peptide (anti-CCP) kit and by enzyme-linked immunosorbent assays (ELISAs) against citrullinated human antigens. Antibody reactivity with P gingivalis was tested by ELISAs against outer membrane antigens (OMAs) and citrullinated enolase from P gingivalis. Results Approximately 19.5{\%} of plasmablast-derived antibodies from anti-CCP-positive RA patients, but none from 1 anti-CCP-negative RA patient or the healthy controls, specifically recognized citrullinated antigens. The immunoglobulin genes encoding these ACPAs were highly mutated, with increased ratios of replacement mutations to silent mutations, suggesting the involvement of active antigen selection in ACPA generation. Interestingly, 63{\%} of the ACPAs cross-reacted with OMAs and/or citrullinated enolase from P gingivalis. The reactivity of ACPAs against citrullinated proteins from P gingivalis was confirmed by immunoblotting and mass spectrometry. Furthermore, some germline-reverted ACPAs retained their reactivity with P gingivalis antigens but completely lost their reactivity with citrullinated human antigens. Conclusion These results suggest that circulating plasmablasts in RA patients produce ACPAs and that this process may be facilitated by anti-P gingivalis immune responses.",
author = "Song Li and Yangsheng Yu and Yinshi Yue and Hongyan Liao and Wanqin Xie and Jessica Thai and Mikuls, {Ted R} and Thiele, {Geoffrey Milton} and Duryee, {Michael J.} and Harlan Sayles and Payne, {Jeffrey B} and Klassen, {Lynell Warren} and O'Dell, {James Robert} and Zhixin Zhang and Kaihong Su",
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T1 - Autoantibodies from Single Circulating Plasmablasts React with Citrullinated Antigens and Porphyromonas gingivalis in Rheumatoid Arthritis

AU - Li, Song

AU - Yu, Yangsheng

AU - Yue, Yinshi

AU - Liao, Hongyan

AU - Xie, Wanqin

AU - Thai, Jessica

AU - Mikuls, Ted R

AU - Thiele, Geoffrey Milton

AU - Duryee, Michael J.

AU - Sayles, Harlan

AU - Payne, Jeffrey B

AU - Klassen, Lynell Warren

AU - O'Dell, James Robert

AU - Zhang, Zhixin

AU - Su, Kaihong

PY - 2016/3/1

Y1 - 2016/3/1

N2 - Objective Anti-citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA). However, the molecular basis for ACPA production is still unclear. The purpose of this study was to determine if circulating plasmablasts from RA patients produce ACPAs and whether Porphyromonas gingivalis facilitates the generation of ACPAs. Methods Using a single-cell antibody cloning approach, we generated 217 and 110 monoclonal recombinant antibodies from circulating plasmablasts from 7 RA patients and 4 healthy controls, respectively. Antibody reactivity with citrullinated antigens was tested by a second-generation anti-cyclic citrullinated peptide (anti-CCP) kit and by enzyme-linked immunosorbent assays (ELISAs) against citrullinated human antigens. Antibody reactivity with P gingivalis was tested by ELISAs against outer membrane antigens (OMAs) and citrullinated enolase from P gingivalis. Results Approximately 19.5% of plasmablast-derived antibodies from anti-CCP-positive RA patients, but none from 1 anti-CCP-negative RA patient or the healthy controls, specifically recognized citrullinated antigens. The immunoglobulin genes encoding these ACPAs were highly mutated, with increased ratios of replacement mutations to silent mutations, suggesting the involvement of active antigen selection in ACPA generation. Interestingly, 63% of the ACPAs cross-reacted with OMAs and/or citrullinated enolase from P gingivalis. The reactivity of ACPAs against citrullinated proteins from P gingivalis was confirmed by immunoblotting and mass spectrometry. Furthermore, some germline-reverted ACPAs retained their reactivity with P gingivalis antigens but completely lost their reactivity with citrullinated human antigens. Conclusion These results suggest that circulating plasmablasts in RA patients produce ACPAs and that this process may be facilitated by anti-P gingivalis immune responses.

AB - Objective Anti-citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA). However, the molecular basis for ACPA production is still unclear. The purpose of this study was to determine if circulating plasmablasts from RA patients produce ACPAs and whether Porphyromonas gingivalis facilitates the generation of ACPAs. Methods Using a single-cell antibody cloning approach, we generated 217 and 110 monoclonal recombinant antibodies from circulating plasmablasts from 7 RA patients and 4 healthy controls, respectively. Antibody reactivity with citrullinated antigens was tested by a second-generation anti-cyclic citrullinated peptide (anti-CCP) kit and by enzyme-linked immunosorbent assays (ELISAs) against citrullinated human antigens. Antibody reactivity with P gingivalis was tested by ELISAs against outer membrane antigens (OMAs) and citrullinated enolase from P gingivalis. Results Approximately 19.5% of plasmablast-derived antibodies from anti-CCP-positive RA patients, but none from 1 anti-CCP-negative RA patient or the healthy controls, specifically recognized citrullinated antigens. The immunoglobulin genes encoding these ACPAs were highly mutated, with increased ratios of replacement mutations to silent mutations, suggesting the involvement of active antigen selection in ACPA generation. Interestingly, 63% of the ACPAs cross-reacted with OMAs and/or citrullinated enolase from P gingivalis. The reactivity of ACPAs against citrullinated proteins from P gingivalis was confirmed by immunoblotting and mass spectrometry. Furthermore, some germline-reverted ACPAs retained their reactivity with P gingivalis antigens but completely lost their reactivity with citrullinated human antigens. Conclusion These results suggest that circulating plasmablasts in RA patients produce ACPAs and that this process may be facilitated by anti-P gingivalis immune responses.

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