{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T16:31:28Z","timestamp":1787761888817,"version":"build-2784847793"},"reference-count":41,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,10,24]],"date-time":"2022-10-24T00:00:00Z","timestamp":1666569600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011892","name":"Kaohsiung Chung-Gung Memorial Hospital Research Funding","doi-asserted-by":"publisher","award":["CRRPG8K0063"],"award-info":[{"award-number":["CRRPG8K0063"]}],"id":[{"id":"10.13039\/501100011892","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>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\u2013protein 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 &gt; 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\u2013protein 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.<\/jats:p>","DOI":"10.3390\/cells11213350","type":"journal-article","created":{"date-parts":[[2022,10,24]],"date-time":"2022-10-24T11:53:55Z","timestamp":1666612435000},"page":"3350","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["A Study on MDA5 Signaling in Splenic B Cells from an Imiquimod-Induced Lupus Mouse Model with Proteomics"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7274-3458","authenticated-orcid":false,"given":"Yu-Jih","family":"Su","sequence":"first","affiliation":[{"name":"Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Departments of Nuclear Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0580-7765","authenticated-orcid":false,"given":"Fu-An","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jim Jinn-Chyuan","family":"Sheu","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung 804201, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3016-9718","authenticated-orcid":false,"given":"Sung-Chou","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Medical Research and Core Laboratory for Phenomics and Diagnostics, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shao-Wen","family":"Weng","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9936-6074","authenticated-orcid":false,"given":"Feng-Chih","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4491-0076","authenticated-orcid":false,"given":"Yen-Hsiang","family":"Chang","sequence":"additional","affiliation":[{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6390-1178","authenticated-orcid":false,"given":"Huan-Yuan","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9003-4065","authenticated-orcid":false,"given":"Chia-Wei","family":"Liou","sequence":"additional","affiliation":[{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Department of Neurology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsu-Kung","family":"Lin","sequence":"additional","affiliation":[{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Department of Neurology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiin-Haur","family":"Chuang","sequence":"additional","affiliation":[{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Pediatric Surgery, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6369-8712","authenticated-orcid":false,"given":"Pei-Wen","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"},{"name":"Center for Mitochondrial Research and Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1002\/art.42021","article-title":"Identification of Shared and Asian-Specific Loci for Systemic Lupus Erythematosus and Evidence for Roles of Type III Interferon Signaling and Lysosomal Function in the Disease: A Multi-Ancestral Genome-Wide Association Study","volume":"74","author":"Wang","year":"2022","journal-title":"Arthritis Rheumatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1186\/s12967-014-0303-1","article-title":"Investigation of the caspase-dependent mitochondrial apoptotic pathway in mononuclear cells of patients with systemic Lupus erythematosus","volume":"12","author":"Su","year":"2014","journal-title":"J. Transl. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/s00281-021-00900-w","article-title":"Recent advances in understanding the genetic basis of systemic lupus erythematosus","volume":"44","author":"Ha","year":"2021","journal-title":"Semin. Immunopathol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1089\/vim.2018.0087","article-title":"Inverse Association Between Antiviral Immunity and Lupus Disease Activity","volume":"31","author":"Su","year":"2018","journal-title":"Viral Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1007\/s00430-007-0040-7","article-title":"Incidental discovery of high systemic lupus erythematosus disease activity associated with cytomegalovirus viral activity","volume":"196","author":"Su","year":"2007","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1007\/s00430-004-0230-5","article-title":"Detecting Epstein-Barr virus DNA from peripheral blood mononuclear cells in adult patients with systemic lupus erythematosus in Taiwan","volume":"194","author":"Yu","year":"2005","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_7","first-page":"44","article-title":"High prevalence of immunoglobulin A antibody against Epstein-Barr virus capsid antigen in adult patients with lupus with disease flare: Case control studies","volume":"32","author":"Chen","year":"2005","journal-title":"J. Rheumatol."},{"key":"ref_8","first-page":"230","article-title":"Overall Status of Epstein-Barr virus Infection, IFN-a, and TLR-7\/9 in Patients with Systemic Lupus Erythematous","volume":"18","author":"Izadi","year":"2021","journal-title":"Iran. J. Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e00987-19","DOI":"10.1128\/JVI.00987-19","article-title":"Endosomal Toll-Like Receptors Mediate Enhancement of Interleukin-17A Production Triggered by Epstein-Barr Virus DNA in Mice","volume":"93","author":"Shehab","year":"2019","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2160","DOI":"10.3389\/fimmu.2019.02160","article-title":"Characterization of cxorf21 Provides Molecular Insight Into Female-Bias Immune Response in SLE Pathogenesis","volume":"10","author":"Harris","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_11","first-page":"126","article-title":"Belimumab slows organ damage progression","volume":"15","author":"McHugh","year":"2019","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1840","DOI":"10.4049\/jimmunol.0902592","article-title":"TLR9 regulates TLR7- and MyD88-dependent autoantibody production and disease in a murine model of lupus","volume":"184","author":"Nickerson","year":"2010","journal-title":"J. Immunol."},{"key":"ref_13","first-page":"13","article-title":"Interferon Induced with Helicase C Domain 1 (IFIH1) Gene single nucleotide polymorphism rs1990760 and its correlation to lupus nephritis: A cross-sectional study","volume":"33","author":"Wang","year":"2019","journal-title":"Formos. J. Rheumatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1002\/emmm.201000120","article-title":"A loss-of-function variant of the antiviral molecule MAVS is associated with a subset of systemic lupus patients","volume":"3","author":"Pothlichet","year":"2011","journal-title":"EMBO Mol. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1111\/j.1365-2249.2010.04176.x","article-title":"Nucleic acid sensing receptors in systemic lupus erythematosus: Development of novel DNA- and\/or RNA-like analogues for treating lupus","volume":"161","author":"Lenert","year":"2010","journal-title":"Clin. Exp. Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"155633","DOI":"10.1016\/j.cyto.2021.155633","article-title":"Abnormalities of the type I interferon signaling pathway in lupus autoimmunity","volume":"146","author":"Gallucci","year":"2021","journal-title":"Cytokine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"897500","DOI":"10.3389\/fimmu.2022.897500","article-title":"A Novel Type I Interferon Primed Dendritic Cell Subpopulation in TREX1 Mutant Chilblain Lupus Patients","volume":"13","author":"Eugster","year":"2022","journal-title":"Front. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1177\/0961203320920706","article-title":"Hsa_circ_0012919 regulates expression of MDA5 by miR-125a-3p in CD4+ T cells of systemic lupus erythematous","volume":"29","author":"Zhang","year":"2020","journal-title":"Lupus"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2452","DOI":"10.3389\/fimmu.2019.02452","article-title":"Antiviral Adaptor MAVS Promotes Murine Lupus with a B Cell Autonomous Role","volume":"10","author":"Sun","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1136\/annrheumdis-2020-218636","article-title":"Molecular pathways in patients with systemic lupus erythematosus revealed by gene-centred DNA sequencing","volume":"80","author":"Sandling","year":"2021","journal-title":"Ann. Rheum. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"657","DOI":"10.2217\/ijr.13.58","article-title":"Genetics of the type I interferon pathway in systemic lupus erythematosus","volume":"8","author":"Niewold","year":"2013","journal-title":"Int. J. Clin. Rheumtol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1861","DOI":"10.1002\/art.20254","article-title":"Induction of interferon-alpha production in plasmacytoid dendritic cells by immune complexes containing nucleic acid released by necrotic or late apoptotic cells and lupus IgG","volume":"50","author":"Lovgren","year":"2004","journal-title":"Arthritis Rheum."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/s13075-017-1501-z","article-title":"Type I IFN signature in childhood-onset systemic lupus erythematosus: A conspiracy of DNA- and RNA-sensing receptors?","volume":"20","author":"Wahadat","year":"2018","journal-title":"Arthritis Res. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"103161","DOI":"10.1016\/j.autrev.2022.103161","article-title":"Friend or foe: RIG- I like receptors and diseases","volume":"21","author":"Song","year":"2022","journal-title":"Autoimmun. Rev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"418","DOI":"10.3389\/fgene.2014.00418","article-title":"Dysregulation of antiviral helicase pathways in systemic lupus erythematosus","volume":"5","author":"Oliveira","year":"2014","journal-title":"Front. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"319","DOI":"10.3389\/fimmu.2013.00319","article-title":"Nucleic Acid sensors and type I interferon production in systemic lupus erythematosus","volume":"4","author":"Shrivastav","year":"2013","journal-title":"Front. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1002\/art.38298","article-title":"Epicutaneous application of toll-like receptor 7 agonists leads to systemic autoimmunity in wild-type mice: A new model of systemic Lupus erythematosus","volume":"66","author":"Yokogawa","year":"2014","journal-title":"Arthritis Rheumatol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4163","DOI":"10.4049\/jimmunol.1502193","article-title":"Efficient Culture of Human Naive and Memory B Cells for Use as APCs","volume":"197","author":"Su","year":"2016","journal-title":"J. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/978-1-60761-175-2_7","article-title":"PANTHER pathway: An ontology-based pathway database coupled with data analysis tools","volume":"563","author":"Mi","year":"2009","journal-title":"Methods Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1714","DOI":"10.1016\/j.celrep.2015.05.032","article-title":"In Vivo RNAi Screening Identifies MDA5 as a Significant Contributor to the Cellular Defense against Influenza A Virus","volume":"11","author":"Benitez","year":"2015","journal-title":"Cell Rep."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Lin, H.Y., Chuang, J.H., Wang, P.W., Lin, T.K., Wu, M.T., Hsu, W.M., and Chuang, H.C. (2020). 5-aza-2\u2019-Deoxycytidine Induces a RIG-I-Related Innate Immune Response by Modulating Mitochondria Stress in Neuroblastoma. Cells, 9.","DOI":"10.3390\/cells9091920"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"954","DOI":"10.1007\/s10875-013-9887-0","article-title":"Distinct autoantibody profiles in systemic lupus erythematosus patients are selectively associated with TLR7 and TLR9 upregulation","volume":"33","author":"Chauhan","year":"2013","journal-title":"J. Clin. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.immuni.2006.07.013","article-title":"Toll-like receptor 7 and TLR9 dictate autoantibody specificity and have opposing inflammatory and regulatory roles in a murine model of lupus","volume":"25","author":"Christensen","year":"2006","journal-title":"Immunity"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Izumi, T., Morioka, Y., Urayama, S.-I., Motooka, D., Tamura, T., Kawagishi, T., Kanai, Y., Kobayashi, T., Ono, C., and Morinaga, A. (2021). DsRNA Sequencing for RNA Virus Surveillance Using Human Clinical Samples. Viruses, 13.","DOI":"10.3390\/v13071310"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/j.jaut.2009.11.001","article-title":"Critical role of TLR7 in the acceleration of systemic lupus erythematosus in TLR9-deficient mice","volume":"34","author":"Wu","year":"2010","journal-title":"J. Autoimmun."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Hanten, J.A., Vasilakos, J.P., Riter, C.L., Neys, L., Lipson, K.E., Alkan, S.S., and Birmachu, W. (2008). Comparison of human B cell activation by TLR7 and TLR9 agonists. BMC Immunol., 9.","DOI":"10.1186\/1471-2172-9-39"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1084\/jem.20050914","article-title":"Nucleic acids of mammalian origin can act as endogenous ligands for Toll-like receptors and may promote systemic lupus erythematosus","volume":"202","author":"Barrat","year":"2005","journal-title":"J. Exp. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3035","DOI":"10.4049\/jimmunol.1402690","article-title":"TLR7- and TLR9-responsive human B cells share phenotypic and genetic characteristics","volume":"194","author":"Simchoni","year":"2015","journal-title":"J. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1093\/intimm\/dxab006","article-title":"Human SLC15A4 is crucial for TLR-mediated type I interferon production and mitochondrial integrity","volume":"33","author":"Kobayashi","year":"2021","journal-title":"Int. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/nrmicro1645","article-title":"Replication of hepatitis C virus","volume":"5","author":"Moradpour","year":"2007","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1038\/nrmicro.2016.87","article-title":"Influenza virus RNA polymerase: Insights into the mechanisms of viral RNA synthesis","volume":"14","author":"Fodor","year":"2016","journal-title":"Nat. Rev. Microbiol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/21\/3350\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:01:52Z","timestamp":1760144512000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/21\/3350"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,24]]},"references-count":41,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["cells11213350"],"URL":"https:\/\/doi.org\/10.3390\/cells11213350","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,24]]}}}