{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T16:00:55Z","timestamp":1793030455096,"version":"4.5.0"},"reference-count":50,"publisher":"BMJ","issue":"4","license":[{"start":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T00:00:00Z","timestamp":1712707200000},"content-version":"unspecified","delay-in-days":9,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJSP2138"],"award-info":[{"award-number":["JPMJSP2138"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["21K08153"],"award-info":[{"award-number":["21K08153"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP23ama121054"],"award-info":[{"award-number":["JP23ama121054"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2024,3,18]]},"published-print":{"date-parts":[[2024,4]]},"abstract":"<jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Cancer immunotherapy including immune checkpoint inhibitors is only effective for a limited population of patients with cancer. Therefore, the development of novel cancer immunotherapy is anticipated. In preliminary studies, we demonstrated that tetracyclines enhanced T-cell responses. Therefore, we herein investigated the efficacy of tetracyclines on antitumor T-cell responses by human peripheral T cells, murine models, and the lung tumor tissues of patients with non-small cell lung cancer (NSCLC), with a focus on signaling pathways in T cells.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>The cytotoxicity of peripheral and lung tumor-infiltrated human T cells against tumor cells was assessed by using bispecific T-cell engager (BiTE) technology (BiTE-assay system). The effects of tetracyclines on T cells in the peripheral blood of healthy donors and the tumor tissues of patients with NSCLC were examined using the BiTE-assay system in comparison with anti-programmed cell death-1 (PD-1) antibody, nivolumab. T-cell signaling molecules were analyzed by flow cytometry, ELISA, and qRT-PCR. To investigate the in vivo antitumor effects of tetracyclines, tetracyclines were administered orally to BALB\/c mice engrafted with murine tumor cell lines, either in the presence or absence of anti-mouse CD8 inhibitors.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>The results obtained revealed that tetracyclines enhanced antitumor T-cell cytotoxicity with the upregulation of granzyme B and increased secretion of interferon-\u03b3 in human peripheral T cells and the lung tumor tissues of patients with NSCLC. The analysis of T-cell signaling showed that CD69 in both CD4<jats:sup>+<\/jats:sup> and CD8<jats:sup>+<\/jats:sup> T cells was upregulated by minocycline. Downstream of T-cell receptor signaling, Zap70 phosphorylation and Nur77 were also upregulated by minocycline in the early phase after T-cell activation. These changes were not observed in T cells treated with anti-PD-1 antibodies under the same conditions. The administration of tetracyclines exhibited antitumor efficacy with the upregulation of CD69 and increases in tumor antigen-specific T cells in murine tumor models. These changes were canceled by the administration of anti-mouse CD8 inhibitors.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>In conclusion, tetracyclines enhanced antitumor T-cell immunity via Zap70 signaling. These results will contribute to the development of novel cancer immunotherapy.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1136\/jitc-2023-008334","type":"journal-article","created":{"date-parts":[[2024,4,15]],"date-time":"2024-04-15T12:15:11Z","timestamp":1713183311000},"page":"e008334","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":23,"title":["Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway"],"prefix":"10.1136","volume":"12","author":[{"given":"Mari","family":"Tone","sequence":"first","affiliation":[{"name":"Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"},{"name":"Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9891-6003","authenticated-orcid":false,"given":"Kota","family":"Iwahori","sequence":"additional","affiliation":[{"name":"Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"},{"name":"Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michinari","family":"Hirata","sequence":"additional","affiliation":[{"name":"Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"},{"name":"Biopharmaceutical Research Division, Shionogi & Co., Ltd, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Azumi","family":"Ueyama","sequence":"additional","affiliation":[{"name":"Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"},{"name":"Biopharmaceutical Research Division, Shionogi & Co., Ltd, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akiyoshi","family":"Tani","sequence":"additional","affiliation":[{"name":"Compound Library Screening Center, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun-Ichi","family":"Haruta","sequence":"additional","affiliation":[{"name":"Lead Explorating Units, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshito","family":"Takeda","sequence":"additional","affiliation":[{"name":"Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasushi","family":"Shintani","sequence":"additional","affiliation":[{"name":"Department of General Thoracic Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Atsushi","family":"Kumanogoh","sequence":"additional","affiliation":[{"name":"Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"},{"name":"Department of Immunopathology, World Premier International Research Center Initiative (WPI), Immunology Frontier Research Center (IFReC), Osaka University, Osaka, Japan"},{"name":"Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka, Japan"},{"name":"Center for Infectious Diseases for Education and Research (CiDER), Osaka University, Osaka, Japan"},{"name":"Japan Agency for Medical Research and Development \u2013 Core Research for Evolutional Science and Technology (AMED\u2013CREST), Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hisashi","family":"Wada","sequence":"additional","affiliation":[{"name":"Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"239","published-online":{"date-parts":[[2024,4,15]]},"reference":[{"key":"2025100314510234000_12.4.e008334.1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1501824"},{"key":"2025100314510234000_12.4.e008334.2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1417636112"},{"key":"2025100314510234000_12.4.e008334.3","doi-asserted-by":"crossref","DOI":"10.1136\/jitc-2021-002954","article-title":"Mitochondrial reactive oxygen species trigger metformin-dependent antitumor immunity via activation of Nrf2\/Mtorc1\/P62 axis in tumor-infiltrating Cd8t lymphocytes","volume":"9","author":"Nishida","year":"2021","journal-title":"J Immunother Cancer"},{"key":"2025100314510234000_12.4.e008334.4","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1620433114"},{"key":"2025100314510234000_12.4.e008334.5","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-18-0095"},{"key":"2025100314510234000_12.4.e008334.6","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1248\/bpb.b21-00806","article-title":"Tetracyclines enhance anti-tumor T-cell responses induced by a Bispecific T-cell Engager","volume":"45","author":"Noguchi","year":"2022","journal-title":"Biol Pharm Bull"},{"key":"2025100314510234000_12.4.e008334.7","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-09-0547"},{"key":"2025100314510234000_12.4.e008334.8","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-18-0713"},{"key":"2025100314510234000_12.4.e008334.9","doi-asserted-by":"crossref","DOI":"10.1080\/2162402X.2015.1062969","article-title":"Expression of inhibitory receptors on Intratumoral T cells modulates the activity of a T cell-Bispecific antibody targeting folate receptor","volume":"5","author":"Schreiner","year":"2016","journal-title":"Oncoimmunology"},{"key":"2025100314510234000_12.4.e008334.10","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-019-39345-5","article-title":"Peripheral T cell cytotoxicity predicts T cell function in the tumor microenvironment","volume":"9","author":"Iwahori","year":"2019","journal-title":"Sci Rep"},{"key":"2025100314510234000_12.4.e008334.11","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-023-41051-2","article-title":"A randomized phase 2 study on Demeclocycline in patients with mild-to-moderate COVID-19","volume":"13","author":"Iwahori","year":"2023","journal-title":"Sci Rep"},{"key":"2025100314510234000_12.4.e008334.12","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-023-35519-4","article-title":"Impact of minocycline on outcomes of EGFR-mutant non-small cell lung cancer patients treated with EGFR-Tkis","volume":"13","author":"Tone","year":"2023","journal-title":"Sci Rep"},{"key":"2025100314510234000_12.4.e008334.13","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2014.156"},{"key":"2025100314510234000_12.4.e008334.14","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-022-09458-5","article-title":"The impact of Ccr8+ regulatory T cells on cytotoxic T cell function in human lung cancer","volume":"12","author":"Haruna","year":"2022","journal-title":"Sci Rep"},{"key":"2025100314510234000_12.4.e008334.15","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.22625"},{"key":"2025100314510234000_12.4.e008334.16","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2594"},{"key":"2025100314510234000_12.4.e008334.17","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1007\/978-1-4939-9454-0_20","article-title":"Supervised machine learning with CITRUS for single cell biomarker discovery","volume":"1989","author":"Polikowsky","year":"2019","journal-title":"Methods Mol Biol"},{"key":"2025100314510234000_12.4.e008334.18","doi-asserted-by":"crossref","DOI":"10.1186\/s12859-018-2486-6","article-title":"iDEP: an integrated web application for differential expression and pathway analysis of RNA-Seq data","volume":"19","author":"Ge","year":"2018","journal-title":"BMC Bioinformatics"},{"key":"2025100314510234000_12.4.e008334.19","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.1434"},{"key":"2025100314510234000_12.4.e008334.20","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1601301"},{"key":"2025100314510234000_12.4.e008334.21","doi-asserted-by":"crossref","first-page":"1506","DOI":"10.1016\/j.chembiol.2018.09.010","article-title":"Tetracyclines modify translation by targeting key human rRNA substructures","volume":"25","author":"Mortison","year":"2018","journal-title":"Cell Chem Biol"},{"key":"2025100314510234000_12.4.e008334.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2011.09.011"},{"key":"2025100314510234000_12.4.e008334.23","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-14-1203"},{"key":"2025100314510234000_12.4.e008334.24","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-14-0190"},{"key":"2025100314510234000_12.4.e008334.25","doi-asserted-by":"crossref","DOI":"10.1186\/s40425-019-0677-y","article-title":"First-in-human phase 1 study of It1208, a defucosylated humanized anti-Cd4 depleting antibody, in patients with advanced solid tumors","volume":"7","author":"Shitara","year":"2019","journal-title":"J Immunother Cancer"},{"key":"2025100314510234000_12.4.e008334.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2004.07.083"},{"key":"2025100314510234000_12.4.e008334.27","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf1292"},{"key":"2025100314510234000_12.4.e008334.28","doi-asserted-by":"publisher","DOI":"10.1038\/S41590-020-0769-3"},{"key":"2025100314510234000_12.4.e008334.29","doi-asserted-by":"crossref","unstructured":"Spakowicz D , Hoyd R , Muniak M , et al . Inferring the role of the microbiome on survival in patients treated with immune checkpoint inhibitors: causal modeling, timing, and classes of concomitant medications. BMC Cancer 2020;20. doi:10.1186\/s12885-020-06882-6","DOI":"10.1186\/s12885-020-06882-6"},{"key":"2025100314510234000_12.4.e008334.30","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1822001116"},{"key":"2025100314510234000_12.4.e008334.31","doi-asserted-by":"crossref","first-page":"1386","DOI":"10.1158\/2326-6066.CIR-22-0041","article-title":"Treg-dominant tumor microenvironment is responsible for hyperprogressive disease after PD-1 blockade therapy","volume":"10","author":"Wakiyama","year":"2022","journal-title":"Cancer Immunol Res"},{"key":"2025100314510234000_12.4.e008334.32","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1038\/s41590-020-0801-7","article-title":"Pd-1","volume":"21","author":"Aksoylar","year":"2020","journal-title":"Nat Immunol"},{"key":"2025100314510234000_12.4.e008334.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.phrs.2012.10.006"},{"key":"2025100314510234000_12.4.e008334.34","doi-asserted-by":"publisher","DOI":"10.1111\/bph.12139"},{"key":"2025100314510234000_12.4.e008334.35","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00594-16"},{"key":"2025100314510234000_12.4.e008334.36","doi-asserted-by":"publisher","DOI":"10.1177\/10454411910020030201"},{"key":"2025100314510234000_12.4.e008334.37","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(97)00605-4"},{"key":"2025100314510234000_12.4.e008334.38","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.23.13496"},{"key":"2025100314510234000_12.4.e008334.39","doi-asserted-by":"publisher","DOI":"10.1016\/0006-2952(92)90381-R"},{"key":"2025100314510234000_12.4.e008334.40","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1111\/j.1365-2249.1995.tb03864.x","article-title":"The influence of tetracyclines on T cell activation","volume":"102","author":"Kloppenburg","year":"1995","journal-title":"Clin Exp Immunol"},{"key":"2025100314510234000_12.4.e008334.41","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.4.934"},{"key":"2025100314510234000_12.4.e008334.42","doi-asserted-by":"publisher","DOI":"10.1086\/651277"},{"key":"2025100314510234000_12.4.e008334.43","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.210518"},{"key":"2025100314510234000_12.4.e008334.44","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.4155\/bio.11.226","article-title":"Analysis of a second-generation tetracycline antibiotic minocycline in human plasma by LC-MS\/MS","volume":"3","author":"Patel","year":"2011","journal-title":"Bioanalysis"},{"key":"2025100314510234000_12.4.e008334.45","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1016\/j.jhep.2023.04.033","article-title":"Nomenclature, diagnosis and management of drug-induced autoimmune-like hepatitis (DI-ALH): an expert opinion meeting report","volume":"79","author":"Andrade","year":"2023","journal-title":"J Hepatol"},{"key":"2025100314510234000_12.4.e008334.46","doi-asserted-by":"publisher","DOI":"10.1001\/archinte.159.5.493"},{"key":"2025100314510234000_12.4.e008334.47","doi-asserted-by":"publisher","DOI":"10.1007\/s10067-013-2245-z"},{"key":"2025100314510234000_12.4.e008334.48","doi-asserted-by":"publisher","DOI":"10.1016\/j.semarthrit.2012.03.006"},{"key":"2025100314510234000_12.4.e008334.49","doi-asserted-by":"crossref","first-page":"1433","DOI":"10.1016\/S0140-6736(12)62163-1","article-title":"Autoimmune hepatitis","volume":"382","author":"Heneghan","year":"2013","journal-title":"The Lancet"},{"key":"2025100314510234000_12.4.e008334.50","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2013.06.016"}],"container-title":["Journal for ImmunoTherapy of Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jitc-2023-008334","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T21:51:11Z","timestamp":1759528271000},"score":1,"resource":{"primary":{"URL":"https:\/\/jitc.bmj.com\/lookup\/doi\/10.1136\/jitc-2023-008334"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,4]]},"references-count":50,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2024,4,15]]},"published-print":{"date-parts":[[2024,4]]}},"alternative-id":["10.1136\/jitc-2023-008334"],"URL":"https:\/\/doi.org\/10.1136\/jitc-2023-008334","relation":{},"ISSN":["2051-1426"],"issn-type":[{"value":"2051-1426","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,4]]}}}