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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MDPI AG
2022-10-01
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| author | Yu-Jih Su Fu-An Li Jim Jinn-Chyuan Sheu Sung-Chou Li Shao-Wen Weng Feng-Chih Shen Yen-Hsiang Chang Huan-Yuan Chen Chia-Wei Liou Tsu-Kung Lin Jiin-Haur Chuang Pei-Wen Wang |
| author_facet | Yu-Jih Su Fu-An Li Jim Jinn-Chyuan Sheu Sung-Chou Li Shao-Wen Weng Feng-Chih Shen Yen-Hsiang Chang Huan-Yuan Chen Chia-Wei Liou Tsu-Kung Lin Jiin-Haur Chuang Pei-Wen Wang |
| author_sort | Yu-Jih Su |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T19:11:07Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-09T19:11:07Z |
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| spelling | doaj.art-24dc77345d854132af08b28d0e0f68882023-11-24T04:07:18ZengMDPI AGCells2073-44092022-10-011121335010.3390/cells11213350A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with ProteomicsYu-Jih Su0Fu-An Li1Jim Jinn-Chyuan Sheu2Sung-Chou Li3Shao-Wen Weng4Feng-Chih Shen5Yen-Hsiang Chang6Huan-Yuan Chen7Chia-Wei Liou8Tsu-Kung Lin9Jiin-Haur Chuang10Pei-Wen Wang11Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanInstitute of Biomedical Sciences, Academia Sinica, Taipei 11529, TaiwanInstitute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung 804201, TaiwanCenter for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanDepartment of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanDepartment of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanCenter for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanInstitute of Biomedical Sciences, Academia Sinica, Taipei 11529, TaiwanCenter for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanCenter for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanCenter for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanDepartment of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, TaiwanIntroduction: 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.https://www.mdpi.com/2073-4409/11/21/3350MDA5lupusB cellsplenocyteproteomics |
| spellingShingle | Yu-Jih Su Fu-An Li Jim Jinn-Chyuan Sheu Sung-Chou Li Shao-Wen Weng Feng-Chih Shen Yen-Hsiang Chang Huan-Yuan Chen Chia-Wei Liou Tsu-Kung Lin Jiin-Haur Chuang Pei-Wen Wang A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics Cells MDA5 lupus B cell splenocyte 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 | MDA5 lupus B cell splenocyte proteomics |
| url | https://www.mdpi.com/2073-4409/11/21/3350 |
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