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Figure 3.: Detection of citrullinated CII in human synovial fluid. (a) Immunoprecipitation of citrullinated CII from synovial fluid. Western blot staining with the antimodified citrulline antibody (AMC) upon appropriate pretreatment of the nitrocellulose membrane and immunostaining with the anticitrullinated CII mAb ACC2 are shown (+, ELISA-positive synovial fluid; −, ELISA-negative synovial fluid [compare with b]). Shown is a representative result of three independently performed experiments. (b) Capture ELISA for detection of citrullinated CII in synovial fluid specimens obtained from patients with OA, Reac A, and RA. Synovial fluid specimens were collected from patients with OA (n = 47), RA (n = 10), and Reac A (n = 15).

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Other Images from "Structure and pathogenicity of antibodies specific for citrullinated collagen type II in experimental arthritis":


Figure 1. ACC mAb antibody specificity in vitro an...

Figure 2. Citrulline-specific antibodies mediate a...

Figure 3. Detection of citrullinated CII in human ...

Figure 4. Molecular structural analysis of the ACC...

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Abstract

Antibodies to citrulline-modified proteins have a high diagnostic value in rheumatoid arthritis (RA). However, their biological role in disease development is still unclear. To obtain insight into this question, a panel of mouse monoclonal antibodies was generated against a major triple helical collagen type II (CII) epitope (position 359–369; ARGLTGRPGDA) with or without arginines modified by citrullination. These antibodies bind cartilage and synovial tissue, and mediate arthritis in mice. Detection of citrullinated CII from RA patients' synovial fluid demonstrates that cartilage-derived CII is indeed citrullinated in vivo. The structure determination of a Fab fragment of one of these antibodies in complex with a citrullinated peptide showed a surprising β-turn conformation of the peptide and provided information on citrulline recognition. Based on these findings, we propose that autoimmunity to CII, leading to the production of antibodies specific for both native and citrullinated CII, is an important pathogenic factor in the development of RA.


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