{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T23:58:17Z","timestamp":1793750297143,"version":"4.6.1"},"reference-count":55,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"824","funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01AG060890"],"award-info":[{"award-number":["R01AG060890"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01DA051889"],"award-info":[{"award-number":["R01DA051889"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01DA055488"],"award-info":[{"award-number":["R01DA055488"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30AI042853"],"award-info":[{"award-number":["P30AI042853"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["T32AI007309"],"award-info":[{"award-number":["T32AI007309"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["F31AI172625"],"award-info":[{"award-number":["F31AI172625"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R21NS126092"],"award-info":[{"award-number":["R21NS126092"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30AI042853"],"award-info":[{"award-number":["P30AI042853"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Transl. Med."],"published-print":{"date-parts":[[2025,11,12]]},"abstract":"<jats:p>To improve existing synthetic RNA-based vaccines, we previously developed a self-amplifying RNA (saRNA)\u2013based vaccine expressing a membrane-anchored (TM) receptor binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein (S-RBD-TM) and demonstrated that a low dose of this saRNA vaccine elicits robust immune responses. Moreover, a recent clinical trial with an saRNA vaccine incorporating 5-methylcytidine (5mC) (saRNA-5mC) has demonstrated reduced vaccine reactogenicity while maintaining robust humoral responses. In this study, we investigate the mechanisms by which 5mC incorporation attenuates adverse effects while maintaining immunogenicity. We found that incorporation of 5mC into the saRNA platform led to prolonged and robust expression of antigen and attenuated induction of type I interferon, a key driver of reactogenicity, specifically in plasmacytoid dendritic cells (pDCs). As a result, saRNA-5mC alleviated excessive innate immune responses in vivo without impairing B cell and T cell responses against the SARS-CoV-2 RBD. Mechanistically, we demonstrated that the detection of unmodified saRNA in pDCs was mediated by a host cytosolic RNA sensor, RIG-I, and this sensing was abolished with 5mC incorporation. In contrast, saRNA-5mC induced robust innate activation in professional antigen-presenting cells, such as macrophages, in a RIG-I\u2013independent manner, highlighting distinct host sensing mechanisms for synthetic RNAs. Our study provides support for the potential clinical use of saRNA-5mC vaccine platforms.<\/jats:p>","DOI":"10.1126\/scitranslmed.adz2276","type":"journal-article","created":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T18:58:19Z","timestamp":1762973899000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":11,"title":["Incorporation of 5-methylcytidine alleviates RIG-I\u2013mediated innate immune responses to a self-amplifying RNA vaccine"],"prefix":"10.1126","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9207-6923","authenticated-orcid":true,"given":"Wataru","family":"Akahata","sequence":"first","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8999-3456","authenticated-orcid":true,"given":"Mai","family":"Komori","sequence":"additional","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3401-9853","authenticated-orcid":true,"given":"Amber L.","family":"Morey","sequence":"additional","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4534-0524","authenticated-orcid":true,"given":"Andr\u00e9s A.","family":"Qui\u00f1ones-Molina","sequence":"additional","affiliation":[{"name":"Department of Virology, Immunology &amp; Microbiology, Boston University Chobanian &amp; Avedisian School of Medicine, Boston, MA 02118, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-3354-7787","authenticated-orcid":true,"given":"James B.","family":"Hood","sequence":"additional","affiliation":[{"name":"Department of Virology, Immunology &amp; Microbiology, Boston University Chobanian &amp; Avedisian School of Medicine, Boston, MA 02118, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2006-2087","authenticated-orcid":true,"given":"Josiane","family":"Fofana","sequence":"additional","affiliation":[{"name":"Department of Virology, Immunology &amp; Microbiology, Boston University Chobanian &amp; Avedisian School of Medicine, Boston, MA 02118, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7768-5965","authenticated-orcid":true,"given":"Luis","family":"Romero","sequence":"additional","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-0300-0739","authenticated-orcid":true,"given":"Elizabeth","family":"Peters","sequence":"additional","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1979-6355","authenticated-orcid":true,"given":"Jonathan D.","family":"Webber","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9172-8200","authenticated-orcid":true,"given":"Tyler","family":"Meeks","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4446-5452","authenticated-orcid":true,"given":"Paulina A.","family":"Przygonska","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-3173-3634","authenticated-orcid":true,"given":"Isabel","family":"Steinberg","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ellison","family":"Ober","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae","family":"Kim","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9766-4166","authenticated-orcid":true,"given":"Daniel C.","family":"Rogan","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7746-3028","authenticated-orcid":true,"given":"Kenta","family":"Matsuda","sequence":"additional","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan F.","family":"Smith","sequence":"additional","affiliation":[{"name":"VLP Therapeutics, Inc., Gaithersburg, MD 20878, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8606-8481","authenticated-orcid":true,"given":"Suryaram","family":"Gummuluru","sequence":"additional","affiliation":[{"name":"Department of Virology, Immunology &amp; Microbiology, Boston University Chobanian &amp; Avedisian School of Medicine, Boston, MA 02118, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2108-8705","authenticated-orcid":true,"given":"Mark","family":"Connors","sequence":"additional","affiliation":[{"name":"HIV-Specific Immunity Section of the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9228-0465","authenticated-orcid":true,"given":"Hisashi","family":"Akiyama","sequence":"additional","affiliation":[{"name":"Department of Virology, Immunology &amp; Microbiology, Boston University Chobanian &amp; Avedisian School of Medicine, Boston, MA 02118, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1080\/14760584.2022.2098719"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1080\/14760584.2023.2208216"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-019-0132-6"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-38457-x"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1997.8878"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41434-020-00204-y"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.12.10265-10269.1998"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.2.507"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(83)90099-5"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M104786200"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M910340199"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(99)00321-X"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.3390\/v10030105"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2008.10.030"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.eclinm.2021.101262"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xcrm.2023.101134"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2005.06.008"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2008.200"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-022-00549-y"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2020.06.011"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2019.01.020"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2024.108964"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05899-7"},{"key":"e_1_3_2_25_2","first-page":"174615","article-title":"Bioactivity determination of native and variant forms of therapeutic interferons","volume":"2011","author":"Larocque L.","year":"2011","unstructured":"L. Larocque, A. Bliu, R. Xu, A. Diress, J. Wang, R. Lin, R. He, M. Girard, X. Li, Bioactivity determination of native and variant forms of therapeutic interferons. J. Biomed. Biotechnol. 2011, 174615 (2011).","journal-title":"J. Biomed. Biotechnol."},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2622-0"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03275-y"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2023.06.017"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(98)00341-7"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2017.08.006"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1038\/nri2056"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(95)90076-4"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2018.12.027"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2020.06.014"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200612056"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.9b00810"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-024-02306-z"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-022-01160-y"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-052521-035330"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2016.05.046"},{"key":"e_1_3_2_41_2","doi-asserted-by":"crossref","unstructured":"C. Ngo K. Rahmani M. Valente N. Collinet G. Bessou M. Fabregue S. Sharkaoui S. Mazzoli C. Pierini-Malosse A. Sansoni F. Fiore C. Laprie M. Gaya C. Gregoire A. Broggi P. Milpied B. Escali\u00e8re T. P. Vu Manh M. Fallet L. Chasson H. Tran M. Le Bert B. Malissen A. Zarubica M. Dalod E. Tomasello Plasmacytoid dendritic cells are dispensable or detrimental in murine systemic or respiratory viral infections. bioRxiv 594961 [Preprint] (2024); https:\/\/doi.org\/10.1101\/2024.05.20.594961.","DOI":"10.1101\/2024.05.20.594961"},{"key":"e_1_3_2_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-020-00602-7"},{"key":"e_1_3_2_43_2","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.aje.a120178"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(89)90347-4"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2016.05.019"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1002\/bit.25170"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1010479"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.03.013"},{"key":"e_1_3_2_49_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.21.11382"},{"key":"e_1_3_2_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00972-17"},{"key":"e_1_3_2_51_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01118-06"},{"key":"e_1_3_2_52_2","doi-asserted-by":"publisher","DOI":"10.1016\/0304-3940(94)11792-H"},{"key":"e_1_3_2_53_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12974-018-1288-0"},{"key":"e_1_3_2_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2021.100351"},{"key":"e_1_3_2_55_2","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"e_1_3_2_56_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2028436"}],"container-title":["Science Translational Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/scitranslmed.adz2276","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T18:58:28Z","timestamp":1762973908000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/scitranslmed.adz2276"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,12]]},"references-count":55,"journal-issue":{"issue":"824","published-print":{"date-parts":[[2025,11,12]]}},"alternative-id":["10.1126\/scitranslmed.adz2276"],"URL":"https:\/\/doi.org\/10.1126\/scitranslmed.adz2276","relation":{},"ISSN":["1946-6234","1946-6242"],"issn-type":[{"value":"1946-6234","type":"print"},{"value":"1946-6242","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,12]]},"assertion":[{"value":"2025-05-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-30","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-12","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"eadz2276"}}