{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:58:33Z","timestamp":1792789113046,"version":"4.5.0"},"reference-count":26,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T00:00:00Z","timestamp":1762992000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Beijing Celarts Biosciences Group Co., Ltd","award":["2021045"],"award-info":[{"award-number":["2021045"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82203683"],"award-info":[{"award-number":["82203683"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82403611"],"award-info":[{"award-number":["82403611"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Toll-like receptor 9 (TLR9) and Toll-like receptor 3 (TLR3), which are widely expressed in dendritic cells (DCs), function as key pattern recognition receptors (PRRs) in the immune system. Their primary roles involve specifically detecting pathogen-associated molecular patterns (PAMPs): TLR9 recognizes unmethylated CpG motifs predominantly found in bacterial and viral DNA, while TLR3 identifies viral double-stranded RNA (dsRNA), a molecular signature associated with viral replication. Their specific agonists [CpG ODN (a TLR9 agonist) and poly(I:C) (a TLR3 agonist)] can effectively activate DCs and enhance the expression of immune activation-related molecules. In this study, by establishing a mouse primary dendritic cell model and a glioma-bearing mouse model, and employing techniques such as transcriptome sequencing, we found that combined stimulation with CpG ODN and poly(I:C) significantly enhanced the anti-tumor function of DCs: in vitro, DCs subjected to combined stimulation showed upregulation of anti-tumor-related surface markers, enhanced migratory capacity, and a more effective activation of CD8+ T cells; in vivo, a DC vaccine loaded with tumor lysate antigen and stimulated with this combined regimen significantly delayed the progression of glioma in tumor-bearing mice. Further investigation revealed that the underlying mechanism for this enhanced effect may involve TLR9 activation promoting TLR3 upregulation through the p38 MAPK-ATF3 signaling axis. Consequently, we designed a sequential stimulation protocol (first CpG ODN then poly(I:C)), which demonstrated a stronger anti-glioma effect compared to simple combined stimulation. This study provides a new strategy for enhancing the immune efficacy of DC vaccines and has potential significance for promoting the clinical translation of DC vaccines.<\/jats:p>","DOI":"10.3390\/cells14221785","type":"journal-article","created":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T09:06:36Z","timestamp":1763111196000},"page":"1785","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["CpG ODN Activates TLR9 and Upregulates TLR3 via the p38 MAPK-ATF3 Signaling Axis to Synergistically Enhance Dendritic Cell Vaccine Efficacy"],"prefix":"10.3390","volume":"14","author":[{"given":"Lv","family":"Zhou","sequence":"first","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Hubei Key Laboratory of Neural Injury and Functional Reconstruction, Huazhong University of Science and Technology, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuowei","family":"Lei","sequence":"additional","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Jiang","sequence":"additional","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Hubei Key Laboratory of Neural Injury and Functional Reconstruction, Huazhong University of Science and Technology, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linpeng","family":"Xu","sequence":"additional","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Hubei Key Laboratory of Neural Injury and Functional Reconstruction, Huazhong University of Science and Technology, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quanji","family":"Wang","sequence":"additional","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Hubei Key Laboratory of Neural Injury and Functional Reconstruction, Huazhong University of Science and Technology, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yimin","family":"Huang","sequence":"additional","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Hubei Key Laboratory of Neural Injury and Functional Reconstruction, Huazhong University of Science and Technology, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3241-7567","authenticated-orcid":false,"given":"Ting","family":"Lei","sequence":"additional","affiliation":[{"name":"Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095, Wuhan 430030, China"},{"name":"Hubei Key Laboratory of Neural Injury and Functional Reconstruction, Huazhong University of Science and Technology, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,13]]},"reference":[{"key":"ref_1","first-page":"1035","article-title":"Identification of KIF15 as a potential therapeutic target and prognostic factor for glioma","volume":"43","author":"Wang","year":"2020","journal-title":"Oncol. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1038\/mt.2012.210","article-title":"T cells redirected to EphA2 for the immunotherapy of glioblastoma","volume":"21","author":"Chow","year":"2013","journal-title":"Mol. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1186\/s13046-020-01567-1","article-title":"Long non-coding RNA NCK1-AS1 promotes the tumorigenesis of glioma through sponging microRNA-138-2-3p and activating the TRIM24\/Wnt\/\u03b2-catenin axis","volume":"39","author":"Huang","year":"2020","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1587","DOI":"10.1158\/0008-5472.CAN-08-2915","article-title":"Effective immunotherapy against murine gliomas using type 1 polarizing dendritic cells\u2013significant roles of CXCL10","volume":"69","author":"Fujita","year":"2009","journal-title":"Cancer Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1038\/s41422-022-00639-5","article-title":"Meningeal lymphatics regulate radiotherapy efficacy through modulating anti-tumor immunity","volume":"32","author":"Zhou","year":"2022","journal-title":"Cell Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1477","DOI":"10.2165\/11538040-000000000-00000","article-title":"Drug or vaccine?: Selecting the appropriate treatment for malignant glioma patients","volume":"70","author":"Dai","year":"2010","journal-title":"Drugs"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"7559","DOI":"10.1158\/0008-5472.CAN-07-0309","article-title":"Nitric oxide boosts chemoimmunotherapy via inhibition of acid sphingomyelinase in a mouse model of melanoma","volume":"67","author":"Perrotta","year":"2007","journal-title":"Cancer Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1742","DOI":"10.1158\/2159-8290.CD-21-0900","article-title":"Vitamin E Enhances Cancer Immunotherapy by Reinvigorating Dendritic Cells via Targeting Checkpoint SHP1","volume":"12","author":"Yuan","year":"2022","journal-title":"Cancer Discov."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1016\/j.tibtech.2020.05.003","article-title":"Rethinking Cancer Immunotherapy by Embracing and Engineering Complexity","volume":"38","author":"Chin","year":"2020","journal-title":"Trends Biotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/s41573-018-0006-z","article-title":"Delivery technologies for cancer immunotherapy","volume":"18","author":"Riley","year":"2019","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e003663","DOI":"10.1136\/jitc-2021-003663","article-title":"Loss of YTHDF1 in gastric tumors restores sensitivity to antitumor immunity by recruiting mature dendritic cells","volume":"10","author":"Bai","year":"2022","journal-title":"J. Immunother. Cancer"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7508928","DOI":"10.1155\/2022\/7508928","article-title":"The Beneficial Effect of IL-12 and IL-18 Transduced Dendritic Cells Stimulated with Tumor Antigens on Generation of an Antitumor Response in a Mouse Colon Carcinoma Model","volume":"2022","author":"Mierzejewska","year":"2022","journal-title":"J. Immunol. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1559325819832144","DOI":"10.1177\/1559325819832144","article-title":"Exposure to Low-Dose Radiation Enhanced the Antitumor Effect of a Dendritic Cell Vaccine","volume":"17","author":"Wang","year":"2019","journal-title":"Dose Response"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1186\/s12967-018-1653-x","article-title":"A lipidated bi-epitope vaccine comprising of MHC-I and MHC-II binder peptides elicits protective CD4 T cell and CD8 T cell immunity against Mycobacterium tuberculosis","volume":"16","author":"Rai","year":"2018","journal-title":"J. Transl. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"9816272","DOI":"10.34133\/2022\/9816272","article-title":"Photothermal Nano-Vaccine Promoting Antigen Presentation and Dendritic Cells Infiltration for Enhanced Immunotherapy of Melanoma via Transdermal Microneedles Delivery","volume":"2022","author":"Zhu","year":"2022","journal-title":"Research"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e004799","DOI":"10.1136\/jitc-2022-004799","article-title":"Intratumoral immunotherapy using a TLR2\/3 agonist, L-pampo, induces robust antitumor immune responses and enhances immune checkpoint blockade","volume":"10","author":"Lee","year":"2022","journal-title":"J. Immunother. Cancer"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e006714","DOI":"10.1136\/jitc-2023-006714","article-title":"Dual TLR9 and PD-L1 targeting unleashes dendritic cells to induce durable antitumor immunity","volume":"11","author":"Cianciaruso","year":"2023","journal-title":"J. Immunother. Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3574","DOI":"10.1158\/0008-5472.CAN-17-3376","article-title":"Sialic Acid Blockade Suppresses Tumor Growth by Enhancing T-cell-Mediated Tumor Immunity","volume":"78","author":"Boltje","year":"2018","journal-title":"Cancer Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2935","DOI":"10.1038\/s41467-021-23244-3","article-title":"PLGA-particle vaccine carrying TLR3\/RIG-I ligand Riboxxim synergizes with immune checkpoint blockade for effective anti-cancer immunotherapy","volume":"12","author":"Koerner","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6428","DOI":"10.1523\/JNEUROSCI.0666-20.2020","article-title":"Synergistic Toll-like Receptor 3\/9 Signaling Affects Properties and Impairs Glioma-Promoting Activity of Microglia","volume":"40","author":"Huang","year":"2020","journal-title":"J. Neurosci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1186\/s13046-020-01673-0","article-title":"CREB5 promotes invasiveness and metastasis in colorectal cancer by directly activating MET","volume":"39","author":"Wang","year":"2020","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1016\/j.cell.2020.06.026","article-title":"Microglia Require CD4 T Cells to Complete the Fetal-to-Adult Transition","volume":"182","author":"Pasciuto","year":"2020","journal-title":"Cell"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1158\/2159-8290.CD-17-1368","article-title":"Chimeric Antigen Receptor T Cell-Mediated Neurotoxicity in Nonhuman Primates","volume":"8","author":"Taraseviciute","year":"2018","journal-title":"Cancer Discov."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Cui, Y., Yu, H., Bu, Z., Wen, L., Yan, L., and Feng, J. (2022). Focus on the Role of the NLRP3 Inflammasome in Multiple Sclerosis: Pathogenesis, Diagnosis, and Therapeutics. Front. Mol. Neurosci., 15.","DOI":"10.3389\/fnmol.2022.894298"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1038\/s41419-023-05692-5","article-title":"Supplementation with \u03b1-ketoglutarate improved the efficacy of anti-PD1 melanoma treatment through epigenetic modulation of PD-L1","volume":"14","author":"Liu","year":"2023","journal-title":"Cell Death Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1038\/s41401-021-00631-6","article-title":"The modulation of PD-L1 induced by the oncogenic HBXIP for breast cancer growth","volume":"43","author":"Xu","year":"2022","journal-title":"Acta Pharmacol. Sin."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/22\/1785\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T09:24:27Z","timestamp":1763112267000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/22\/1785"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,13]]},"references-count":26,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["cells14221785"],"URL":"https:\/\/doi.org\/10.3390\/cells14221785","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,13]]}}}