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A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics

Introduction: Several environmental stimuli may influence lupus, particularly viral infections. In this study, we used an imiquimod-induced lupus mouse model focused on the TLR7 pathway and proteomics analysis to determine the specific pathway related to viral infection and the related protein expre...

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Autores principales: Su, Yu-Jih, Li, Fu-An, Sheu, Jim Jinn-Chyuan, Li, Sung-Chou, Weng, Shao-Wen, Shen, Feng-Chih, Chang, Yen-Hsiang, Chen, Huan-Yuan, Liou, Chia-Wei, Lin, Tsu-Kung, Chuang, Jiin-Haur, Wang, Pei-Wen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9657434/
https://www.ncbi.nlm.nih.gov/pubmed/36359746
http://dx.doi.org/10.3390/cells11213350
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author Su, Yu-Jih
Li, Fu-An
Sheu, Jim Jinn-Chyuan
Li, Sung-Chou
Weng, Shao-Wen
Shen, Feng-Chih
Chang, Yen-Hsiang
Chen, Huan-Yuan
Liou, Chia-Wei
Lin, Tsu-Kung
Chuang, Jiin-Haur
Wang, Pei-Wen
author_facet Su, Yu-Jih
Li, Fu-An
Sheu, Jim Jinn-Chyuan
Li, Sung-Chou
Weng, Shao-Wen
Shen, Feng-Chih
Chang, Yen-Hsiang
Chen, Huan-Yuan
Liou, Chia-Wei
Lin, Tsu-Kung
Chuang, Jiin-Haur
Wang, Pei-Wen
author_sort Su, Yu-Jih
collection PubMed
description Introduction: Several environmental stimuli may influence lupus, particularly viral infections. In this study, we used an imiquimod-induced lupus mouse model focused on the TLR7 pathway and proteomics analysis to determine the specific pathway related to viral infection and the related protein expressions in splenic B cells to obtain insight into B-cell responses to viral infection in the lupus model. Materials and Methods: We treated FVB/N wild-type mice with imiquimod for 8 weeks to induce lupus symptoms and signs, retrieved splenocytes, selected B cells, and conducted the proteomic analysis. The B cells were co-cultured with CD40L+ feeder cells for another week before performing Western blot analysis. Panther pathway analysis was used to disclose the pathways activated and the protein–protein interactome was analyzed by the STRING database in this lupus murine model. Results: The lupus model was well established and well demonstrated with serology evidence and pathology proof of lupus-mimicking organ damage. Proteomics data of splenic B cells revealed that the most important activated pathways (fold enrichment > 100) demonstrated positive regulation of the MDA5 signaling pathway, negative regulation of IP-10 production, negative regulation of chemokine (C-X-C motif) ligand 2 production, and positive regulation of the RIG-I signaling pathway. A unique protein–protein interactome containing 10 genes was discovered, within which ISG15, IFIH1, IFIT1, DDX60, and DHX58 were demonstrated to be downstream effectors of MDA5 signaling. Finally, we found B-cell intracellular cytosolic proteins via Western blot experiment and continued to observe MDA5-related pathway activation. Conclusion: In this experiment, we confirmed that the B cells in the lupus murine model focusing on the TLR7 pathway were activated through the MDA5 signaling pathway, an important RNA sensor implicated in the detection of viral infections and autoimmunity. The MDA5 agonist/antagonist RNAs and the detailed molecular interactions within B cells are worthy of further investigation for lupus therapy.
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spelling pubmed-96574342022-11-15 A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics Su, Yu-Jih Li, Fu-An Sheu, Jim Jinn-Chyuan Li, Sung-Chou Weng, Shao-Wen Shen, Feng-Chih Chang, Yen-Hsiang Chen, Huan-Yuan Liou, Chia-Wei Lin, Tsu-Kung Chuang, Jiin-Haur Wang, Pei-Wen Cells Article Introduction: Several environmental stimuli may influence lupus, particularly viral infections. In this study, we used an imiquimod-induced lupus mouse model focused on the TLR7 pathway and proteomics analysis to determine the specific pathway related to viral infection and the related protein expressions in splenic B cells to obtain insight into B-cell responses to viral infection in the lupus model. Materials and Methods: We treated FVB/N wild-type mice with imiquimod for 8 weeks to induce lupus symptoms and signs, retrieved splenocytes, selected B cells, and conducted the proteomic analysis. The B cells were co-cultured with CD40L+ feeder cells for another week before performing Western blot analysis. Panther pathway analysis was used to disclose the pathways activated and the protein–protein interactome was analyzed by the STRING database in this lupus murine model. Results: The lupus model was well established and well demonstrated with serology evidence and pathology proof of lupus-mimicking organ damage. Proteomics data of splenic B cells revealed that the most important activated pathways (fold enrichment > 100) demonstrated positive regulation of the MDA5 signaling pathway, negative regulation of IP-10 production, negative regulation of chemokine (C-X-C motif) ligand 2 production, and positive regulation of the RIG-I signaling pathway. A unique protein–protein interactome containing 10 genes was discovered, within which ISG15, IFIH1, IFIT1, DDX60, and DHX58 were demonstrated to be downstream effectors of MDA5 signaling. Finally, we found B-cell intracellular cytosolic proteins via Western blot experiment and continued to observe MDA5-related pathway activation. Conclusion: In this experiment, we confirmed that the B cells in the lupus murine model focusing on the TLR7 pathway were activated through the MDA5 signaling pathway, an important RNA sensor implicated in the detection of viral infections and autoimmunity. The MDA5 agonist/antagonist RNAs and the detailed molecular interactions within B cells are worthy of further investigation for lupus therapy. MDPI 2022-10-24 /pmc/articles/PMC9657434/ /pubmed/36359746 http://dx.doi.org/10.3390/cells11213350 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Su, Yu-Jih
Li, Fu-An
Sheu, Jim Jinn-Chyuan
Li, Sung-Chou
Weng, Shao-Wen
Shen, Feng-Chih
Chang, Yen-Hsiang
Chen, Huan-Yuan
Liou, Chia-Wei
Lin, Tsu-Kung
Chuang, Jiin-Haur
Wang, Pei-Wen
A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics
title A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics
title_full A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics
title_fullStr A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics
title_full_unstemmed A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics
title_short A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics
title_sort study on mda5 signaling in splenic b cells from an imiquimod-induced lupus mouse model with proteomics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9657434/
https://www.ncbi.nlm.nih.gov/pubmed/36359746
http://dx.doi.org/10.3390/cells11213350
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