Anti-carbamylated protein antibodies drive AEC II toward a profibrotic phenotype by interacting with carbamylated TLR5.
Autor: | Xu W; Department of Rheumatology and Immunology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China., Huang M; Department of Respiratory Medicine, Shandong Provincial Third Hospital, Shandong University, Jinan, China., Dong R; Department of Rheumatology and Immunology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China., Yan S; Department of Rheumatology and Immunology, Shandong Province Hospital of Shandong First Medical University, Jinan, China., An Y; Department of Rheumatology and Immunology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China., Liu B; Department of Rheumatology and Immunology, Shandong Province Hospital of Shandong First Medical University, Jinan, China., Ma Z; Department of Rheumatology and Immunology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.; Department of Rheumatology and Immunology, Shandong Province Hospital of Shandong First Medical University, Jinan, China.; Department of Integrated traditional Chinese and Western Medicine, Shandong University of Traditional Chinese Medicine, Jinan, China., Mu K; Department of Pathology, School of Basic Medical Sciences, Shandong University, Jinan, China.; Department of Pathology, Qilu Hospital of Shandong University, Jinan, China., Yang Q; Department of Rheumatology and Immunology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.; Department of Rheumatology and Immunology, Shandong Province Hospital of Shandong First Medical University, Jinan, China. |
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Jazyk: | angličtina |
Zdroj: | Rheumatology (Oxford, England) [Rheumatology (Oxford)] 2024 Oct 01; Vol. 63 (10), pp. 2874-2886. |
DOI: | 10.1093/rheumatology/keae111 |
Abstrakt: | Objectives: This study looked at the role of anti-carbamylated protein (anti-CarP) antibodies in contributing to lung fibrosis in CTD-associated interstitial lung disease (ILD) in an autoantigen-dependent manner. Methods: ELISA was used to test serum samples, including 89 from the CTD-ILD group and 170 from the non-CTD-ILD group, for anti-CarP levels. Male C57BL/6 mice were used for the pulmonary fibrosis model and anti-CarP treatment in vivo (n = 5) and patient serum-derived or commercialized anti-CarP was used for cell treatment. We identified the carbamylated membrane protein via immunofluorescence (IF) and co-immunoprecipitation followed by mass spectrometry (MS) analysis. Quantitative RT-PCR, IF and western blot were performed to explore the antigen-dependent role of anti-CarP. A native electrophoretic mobility shift assay and MS analysis were used to verify direct interaction and carbamylation sites. Results: A significantly higher serum anti-CarP level was observed in CTD with ILD than without ILD. In vivo, intrapulmonary delivery of anti-CarP induces epithelial-mesenchymal transition (EMT) and microfibrotic foci. Carbamylation was enriched in type II alveolar epithelial cells (AEC II). A novel carbamylated membrane receptor, specifically recognized by anti-CarP, was identified as toll-like receptor 5 (TLR5). We found anti-CarP induces the nuclear translocation of NF-κB and downstream events, including EMT and expression of inflammatory cytokines in AEC II, which were reversed by TLR5 blocking or TLR5 knockdown. Moreover, up to 12 lysine carbamylation sites were found in TLR5 ectodomain, allowing the interaction of anti-CarP with carbamylated TLR5. Conclusions: Overall, we found anti-CarP drives aberrant AEC II activation by interacting with carbamylated TLR5 to promote ILD progression. (© The Author(s) 2024. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.com.) |
Databáze: | MEDLINE |
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