Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis
IntroductionPeptidylarginine deiminases (PADs) mediate citrullination, an irreversible posttranslational modification that converts arginine to citrulline residues in proteins. Rheumatoid arthritis (RA) is characterized by unique autoantibodies that recognize citrullinated peptides, which are highly...
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| Format: | Article |
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Frontiers Media S.A.
2023-01-01
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| Serija: | Frontiers in Immunology |
| Teme: | |
| Online dostop: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1111465/full |
| _version_ | 1828051229934092288 |
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| author | Karen Yu Luna Dillemans Mieke Gouwy Helena Bessa Mieke Metzemaekers Erik Martens Patrick Matthys Xavier Bossuyt Patrick Verschueren Carine Wouters Lien De Somer Paul Proost |
| author_facet | Karen Yu Luna Dillemans Mieke Gouwy Helena Bessa Mieke Metzemaekers Erik Martens Patrick Matthys Xavier Bossuyt Patrick Verschueren Carine Wouters Lien De Somer Paul Proost |
| author_sort | Karen Yu |
| collection | DOAJ |
| description | IntroductionPeptidylarginine deiminases (PADs) mediate citrullination, an irreversible posttranslational modification that converts arginine to citrulline residues in proteins. Rheumatoid arthritis (RA) is characterized by unique autoantibodies that recognize citrullinated peptides, which are highly specific for this disease. However, the mechanism preceding the anti-citrulline response remains largely unclear. PAD enzymes are known to fuel the autoimmune response by generating autoreactive epitopes, and sustain local synovial inflammation through neutrophil extracellular trap formation. Therefore, detecting endogenous PAD activity is important to understand the pathogenesis of arthritis.MethodsIn this study, we improved a fluorescent in vitro assay to enable endogenous PAD activity characterization in complex samples. We combine the use of an in-house synthetic, arginine-rich substrate and a negatively charged dye molecule to visualize enzyme activity.ResultsThis pioneering PAD assay allowed profiling of active citrullination in leukocytes and in local and systemic samples of an arthritis cohort. Our results reveal that RA and juvenile idiopathic arthritis (JIA) synovial fluids display similar levels of PAD activity. In contrast, citrullination was limited in joints of patients suffering from gout or Lyme’s disease. Interestingly, in blood, a higher level of extracellular citrullination was only found in anti-CCP-positive RA patients.DiscussionOur finding suggests that enhanced synovial PAD activity drives the loss in tolerance towards citrullinated proteins and that systemic citrullination may indicate the risk for developing citrulline-specific autoimmunity. |
| first_indexed | 2024-04-10T19:34:39Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-10T19:34:39Z |
| publishDate | 2023-01-01 |
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| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-9ca117eb2c8840c6a6bb1c642c7ab3772023-01-30T08:56:07ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-01-011410.3389/fimmu.2023.11114651111465Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritisKaren Yu0Luna Dillemans1Mieke Gouwy2Helena Bessa3Mieke Metzemaekers4Erik Martens5Patrick Matthys6Xavier Bossuyt7Patrick Verschueren8Carine Wouters9Lien De Somer10Paul Proost11Laboratory of Molecular Immunology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Molecular Immunology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Molecular Immunology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Molecular Immunology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Molecular Immunology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Immunobiology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Immunobiology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory Medicine, University Hospitals Leuven, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumSkeletal Biology and Engineering Research Center, Department of Development and Regeneration, KU Leuven, Leuven, BelgiumLaboratory of Immunobiology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Immunobiology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumLaboratory of Molecular Immunology, Rega Institute for Medical Research, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, BelgiumIntroductionPeptidylarginine deiminases (PADs) mediate citrullination, an irreversible posttranslational modification that converts arginine to citrulline residues in proteins. Rheumatoid arthritis (RA) is characterized by unique autoantibodies that recognize citrullinated peptides, which are highly specific for this disease. However, the mechanism preceding the anti-citrulline response remains largely unclear. PAD enzymes are known to fuel the autoimmune response by generating autoreactive epitopes, and sustain local synovial inflammation through neutrophil extracellular trap formation. Therefore, detecting endogenous PAD activity is important to understand the pathogenesis of arthritis.MethodsIn this study, we improved a fluorescent in vitro assay to enable endogenous PAD activity characterization in complex samples. We combine the use of an in-house synthetic, arginine-rich substrate and a negatively charged dye molecule to visualize enzyme activity.ResultsThis pioneering PAD assay allowed profiling of active citrullination in leukocytes and in local and systemic samples of an arthritis cohort. Our results reveal that RA and juvenile idiopathic arthritis (JIA) synovial fluids display similar levels of PAD activity. In contrast, citrullination was limited in joints of patients suffering from gout or Lyme’s disease. Interestingly, in blood, a higher level of extracellular citrullination was only found in anti-CCP-positive RA patients.DiscussionOur finding suggests that enhanced synovial PAD activity drives the loss in tolerance towards citrullinated proteins and that systemic citrullination may indicate the risk for developing citrulline-specific autoimmunity.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1111465/fullcitrullinationinflammationjoint inflammationpeptidylarginine deiminaseautoimmunity |
| spellingShingle | Karen Yu Luna Dillemans Mieke Gouwy Helena Bessa Mieke Metzemaekers Erik Martens Patrick Matthys Xavier Bossuyt Patrick Verschueren Carine Wouters Lien De Somer Paul Proost Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis Frontiers in Immunology citrullination inflammation joint inflammation peptidylarginine deiminase autoimmunity |
| title | Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis |
| title_full | Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis |
| title_fullStr | Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis |
| title_full_unstemmed | Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis |
| title_short | Novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis |
| title_sort | novel method to quantify peptidylarginine deiminase activity shows distinct citrullination patterns in rheumatoid and juvenile idiopathic arthritis |
| topic | citrullination inflammation joint inflammation peptidylarginine deiminase autoimmunity |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1111465/full |
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