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Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis
Kristine A. Kuhn, … , William H. Robinson, V. Michael Holers
Kristine A. Kuhn, … , William H. Robinson, V. Michael Holers
Published April 3, 2006
Citation Information: J Clin Invest. 2006;116(4):961-973. https://doi.org/10.1172/JCI25422.
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Research Article Autoimmunity

Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis

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Abstract

Antibodies against citrullinated proteins are specific and predictive markers for rheumatoid arthritis although the pathologic relevance of these antibodies remains unclear. To investigate the significance of these autoantibodies, collagen-induced arthritis (CIA) in mice was used to establish an animal model of antibody reactivity to citrullinated proteins. DBA/1J mice were immunized with bovine type II collagen (CII) at days 0 and 21, and serum was collected every 7 days for analysis. Antibodies against both CII and cyclic citrullinated peptide, one such citrullinated antigen, appeared early after immunization, before joint swelling was observed. Further, these antibodies demonstrated specific binding to citrullinated filaggrin in rat esophagus by indirect immunofluorescence and citrullinated fibrinogen by Western blot. To evaluate the role of immune responses to citrullinated proteins in CIA, mice were tolerized with a citrulline-containing peptide, followed by antigen challenge with CII. Tolerized mice demonstrated significantly reduced disease severity and incidence compared with controls. We also identified novel murine monoclonal antibodies specific to citrullinated fibrinogen that enhanced arthritis when coadministered with a submaximal dose of anti-CII antibodies and bound targets within the inflamed synovium of mice with CIA. These results demonstrate that antibodies against citrullinated proteins are centrally involved in the pathogenesis of autoimmune arthritis.

Authors

Kristine A. Kuhn, Liudmila Kulik, Beren Tomooka, Kristin J. Braschler, William P. Arend, William H. Robinson, V. Michael Holers

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Figure 4

Antibodies specific to citrullinated fibrinogen substantially enhance submaximal arthritis.

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Antibodies specific to citrullinated fibrinogen substantially enhance su...
(A) Purified D513, 1042, or 1618 monoclonal antibody was applied to a Western blot of unmodified and citrullinated fibrinogen. To compete for antigen binding on the blot, soluble unmodified or citrullinated fibrinogen was added to D513 prior to applying the antibody to the Western blot. (B) The relative reactivities of the monoclonal antibodies for unmodified and citrullinated fibrinogen were determined by applying increasing amounts of biotinylated unmodified and citrullinated fibrinogen (fibr) and CII to ELISA plates coated with monoclonal antibody. The amount of protein binding was detected as described in Methods and is displayed as OD. (C) Additional antigens recognized by D513, 1042, and 1618 were detected using synovial antigen arrays. The relative reactivity for the antigens recognized is shown. (D) Arthrogen, a cocktail of 4 monoclonal antibodies against CII, was transferred intravenously into 6- to 8-week-old male DBA/1J mice at increasing doses followed by 50 μg LPS intraperitoneally 3 days later in order to determine which dose would establish submaximal disease. n = 3 per dose group. (E) D513 (n = 9) or control monoclonal antibody (n = 6) were administered alone or in combination with the submaximal dose of Arthrogen (1 mg). (F and G) Either 1 mg or 2 mg of monoclonal antibody 1042 or 1618 (n = 6) or 2 mg HB5 control monoclonal antibody (n = 6) was combined with 1 mg of Arthrogen and transferred into mice as performed previously with D513. Data shown are the average disease score per group α SEM. Statistical significance was determined using 1-way ANOVA. *P < 0.05; **P < 0.01 in comparison with submaximal Arrogen alone.

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ISSN: 0021-9738 (print), 1558-8238 (online)

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