{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T10:36:01Z","timestamp":1785321361229,"version":"3.55.0"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T00:00:00Z","timestamp":1772064000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      CD19-directed chimeric antigen receptor (CD19-CAR) T-cell therapy has markedly improved outcomes in relapsed and refractory B-cell malignancies, but its efficacy remains limited by insufficient\n                      <jats:italic>in vivo<\/jats:italic>\n                      persistence and functional exhaustion. We have generated functionally rejuvenated T-cells (rejTs) by reprogramming antigen-specific cytotoxic T lymphocytes (CTLs) into induced pluripotent stem cells (iPSCs) and redifferentiating them into CTLs with restored proliferative capacity. In this study, we explored a vaccine synergy strategy to enhance the persistence of CAR-rejuvenated CTLs (CARrejTs) through T-cell receptor (TCR) restimulation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      SARS-CoV-2 spike protein-specific rejTs (COVID19-rejTs) were established from iPSCs derived from spike protein-specific CTLs. A CD19-CAR was introduced into these iPSCs to generate dual-antigen recognition CARrejTs targeting CD19 and COVID-19 spike protein (1919-CARrejTs). Subsequently, 1919-CARrejTs were assessed for cytotoxicity, proliferative capacity, and exhaustion phenotype using\n                      <jats:sup>51<\/jats:sup>\n                      Cr release assays, sequential rechallenge assays, and CFSE-based proliferation analysis with CAR- or TCR-dependent stimulation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>1919-CARrejTs uniformly expressed both CD19-CAR and spike protein-specific TCRs, retained antigen-specific cytotoxicity, and exhibited a rejuvenated phenotype with higher expression of granzyme B and perforin and lower expression of exhaustion markers compared with conventional CD19-CAR-T cells. Dual-antigen recognition enhanced cytotoxicity under matched antigen presentation, and 1919-CARrejTs maintained durable tumor control in sequential rechallenge assays. CFSE dilution analysis revealed that TCR-mediated stimulation by spike protein-specific peptide provided strong proliferative capacity of 1919-CARrejTs in an HLA-dependent manner.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The combination of iPSC-mediated rejuvenation and dual-antigen recognition via CAR and native TCR confers superior cytotoxicity, persistence, and proliferative potential compared to conventional CD19-CAR-T cells. These findings provide a proof-of-concept for a vaccine-synergy strategy in which\n                      <jats:italic>in vivo<\/jats:italic>\n                      TCR restimulation supports selective expansion and sustained antitumor effect of dual-antigen recognition T-cells that can be a promising treatment approach for B-cell malignancies.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcell.2026.1772146","type":"journal-article","created":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T06:56:25Z","timestamp":1772088985000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Dual-antigen recognition iPSC-derived CAR-T cells for B-cell malignancies: establishment of a COVID-19 vaccine synergy strategy"],"prefix":"10.3389","volume":"14","author":[{"given":"Norihide","family":"Izumi","sequence":"first","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshiki","family":"Furukawa","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shintaro","family":"Kinoshita","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Midori","family":"Ishii","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayaka","family":"Goto","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hajime","family":"Yasuda","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Ando","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]},{"name":"Division of Cell Therapy and Blood Transfusion Medicine, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiromitsu","family":"Nakauchi","sequence":"additional","affiliation":[{"name":"Stem Cell Therapy Division, Advanced Research Initiative, Institute of Science Tokyo","place":["Tokyo, Japan"]},{"name":"Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine","place":["Stanford, CA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miki","family":"Ando","sequence":"additional","affiliation":[{"name":"Department of Hematology, Juntendo University School of Medicine","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,2,26]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1016\/j.exphem.2016.10.009","article-title":"'Off-the-shelf' immunotherapy with iPSC-derived rejuvenated cytotoxic T lymphocytes","volume":"47","author":"Ando","year":"2017","journal-title":"Exp. Hematol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"472","DOI":"10.1038\/cgt.2014.53","article-title":"Bortezomib sensitizes non-small cell lung cancer to mesenchymal stromal cell-delivered inducible caspase-9-mediated cytotoxicity","volume":"21","author":"Ando","year":"2014","journal-title":"Cancer Gene Ther."},{"key":"B3","doi-asserted-by":"publisher","first-page":"597","DOI":"10.1016\/j.stemcr.2015.07.011","article-title":"A safeguard system for induced pluripotent stem cell-derived rejuvenated T cell therapy","volume":"5","author":"Ando","year":"2015","journal-title":"Stem Cell Rep."},{"key":"B4","doi-asserted-by":"publisher","first-page":"796","DOI":"10.3324\/haematol.2019.223511","article-title":"Long-term eradication of extranodal natural killer\/T-cell lymphoma, nasal type, by induced pluripotent stem cell-derived Epstein-Barr virus-specific rejuvenated T cells in vivo","volume":"105","author":"Ando","year":"2020","journal-title":"Haematologica"},{"key":"B5","doi-asserted-by":"publisher","first-page":"113887","DOI":"10.1016\/j.celrep.2024.113887","article-title":"CD8(+) T cell memory induced by successive SARS-CoV-2 mRNA vaccinations is characterized by shifts in clonal dominance","volume":"43","author":"Aoki","year":"2024","journal-title":"Cell Rep."},{"key":"B6","doi-asserted-by":"publisher","first-page":"1524","DOI":"10.1093\/cid\/ciae420","article-title":"Safety and immunogenicity of an mRNA-1273 booster in children","volume":"79","author":"Berthaud","year":"2024","journal-title":"Clin. Infect. Dis."},{"key":"B7","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1038\/s41571-023-00754-1","article-title":"Long-term outcomes following CAR T cell therapy: what we know so far","volume":"20","author":"Cappell","year":"2023","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"2952","DOI":"10.1158\/1078-0432.ccr-14-2998","article-title":"K562-Derived whole-cell vaccine enhances antitumor responses of CAR-Redirected virus-specific cytotoxic T lymphocytes in vivo","volume":"21","author":"Caruana","year":"2015","journal-title":"Clin. Cancer Res."},{"key":"B9","doi-asserted-by":"publisher","first-page":"eabn2231","DOI":"10.1126\/scitranslmed.abn2231","article-title":"Oncolytic virus\u2013mediated expansion of dual-specific CAR T cells improves efficacy against solid tumors in mice","volume":"14","author":"Evgin","year":"2022","journal-title":"Sci. Transl. Med."},{"key":"B10","doi-asserted-by":"publisher","first-page":"101327","DOI":"10.1016\/j.xcrm.2023.101327","article-title":"iPSC-derived hypoimmunogenic tissue resident memory T cells mediate robust anti-tumor activity against cervical cancer","volume":"4","author":"Furukawa","year":"2023","journal-title":"Cell Rep. Med."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1622","DOI":"10.1038\/mt.2012.130","article-title":"Rapidly generated multivirus-specific cytotoxic T lymphocytes for the prophylaxis and treatment of viral infections","volume":"20","author":"Gerdemann","year":"2012","journal-title":"Mol. Ther."},{"key":"B12","doi-asserted-by":"publisher","first-page":"534","DOI":"10.1016\/j.ymthe.2021.10.006","article-title":"Dual-antigen targeted iPSC-derived chimeric antigen receptor-T cell therapy for refractory lymphoma","volume":"30","author":"Harada","year":"2022","journal-title":"Mol. Ther."},{"key":"B13","doi-asserted-by":"publisher","first-page":"95","DOI":"10.2147\/btt.s291768","article-title":"Armored CAR T-Cells: the next chapter in T-Cell cancer immunotherapy","volume":"15","author":"Hawkins","year":"2021","journal-title":"Biol. Targets Ther."},{"key":"B14","doi-asserted-by":"publisher","first-page":"3165","DOI":"10.1182\/blood.2021011568","article-title":"Efficacy of the BNT162b2 mRNA COVID-19 vaccine in patients with chronic lymphocytic leukemia","volume":"137","author":"Herishanu","year":"2021","journal-title":"Blood"},{"key":"B15","doi-asserted-by":"publisher","first-page":"2394","DOI":"10.1016\/j.ymthe.2020.07.004","article-title":"Sustainable tumor-suppressive effect of iPSC-Derived rejuvenated T cells targeting cervical cancers","volume":"28","author":"Honda","year":"2020","journal-title":"Mol. Ther."},{"key":"B16","doi-asserted-by":"publisher","first-page":"112663","DOI":"10.1016\/j.celrep.2023.112663","article-title":"NFAT and NF-\u03baB dynamically co-regulate TCR and CAR signaling responses in human T cells","volume":"42","author":"Huang","year":"2023","journal-title":"Cell Rep."},{"key":"B17","doi-asserted-by":"publisher","first-page":"430","DOI":"10.1038\/s41467-020-20658-3","article-title":"A clinically applicable and scalable method to regenerate T-cells from iPSCs for off-the-shelf T-cell immunotherapy","volume":"12","author":"Iriguchi","year":"2021","journal-title":"Nat. Commun."},{"key":"B18","doi-asserted-by":"publisher","first-page":"2905","DOI":"10.1182\/blood.2020008488","article-title":"SARS-CoV-2\u2013specific T cells are rapidly expanded for therapeutic use and target conserved regions of the membrane protein","volume":"136","author":"Keller","year":"2020","journal-title":"Blood"},{"key":"B19","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1038\/nbt.2459","article-title":"Combinatorial antigen recognition with balanced signaling promotes selective tumor eradication by engineered T cells","volume":"31","author":"Kloss","year":"2013","journal-title":"Nat. Biotechnol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1186\/s12967-022-03442-3","article-title":"Exhaustion of CAR T cells: potential causes and solutions","volume":"20","author":"Kouro","year":"2022","journal-title":"J. Transl. Med."},{"key":"B21","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1016\/j.tips.2024.12.004","article-title":"Boosting CAR-T cell therapy through vaccine synergy","volume":"46","author":"Li","year":"2025","journal-title":"Trends Pharmacol. Sci."},{"key":"B22","doi-asserted-by":"publisher","first-page":"332","DOI":"10.1016\/j.cmi.2022.10.030","article-title":"B-cell malignancies and COVID-19: a narrative review","volume":"29","author":"Luque-Paz","year":"2023","journal-title":"Clin. Microbiol. Infect."},{"key":"B23","doi-asserted-by":"publisher","first-page":"2307","DOI":"10.1182\/blood.2022018893","article-title":"5-Year Follow-Up supports curative potential of axicabtagene Ciloleucel in refractory large B-Cell lymphoma (ZUMA-1)","volume":"141","author":"Neelapu","year":"2023","journal-title":"Blood"},{"key":"B24","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1016\/j.stem.2012.11.002","article-title":"Generation of rejuvenated antigen-specific T cells by reprogramming to pluripotency and redifferentiation","volume":"12","author":"Nishimura","year":"2013","journal-title":"Cell Stem Cell"},{"key":"B25","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1038\/s41586-021-03841-4","article-title":"Rapid and stable mobilization of CD8+ T cells by SARS-CoV-2 mRNA vaccine","volume":"597","author":"Oberhardt","year":"2021","journal-title":"Nature"},{"key":"B26","doi-asserted-by":"publisher","first-page":"2603","DOI":"10.1056\/nejmoa2034577","article-title":"Safety and efficacy of the BNT162b2 mRNA Covid-19 vaccine","volume":"383","author":"Polack","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"B27","doi-asserted-by":"publisher","first-page":"2799","DOI":"10.3324\/haematol.2024.287136","article-title":"T-cell and antibody responses in immunocompromised patients with hematologic malignancies indicate strong potential of SARS-CoV-2 mRNA vaccines","volume":"110","author":"Pothast","year":"2025","journal-title":"Haematologica"},{"key":"B28","doi-asserted-by":"publisher","first-page":"eaaj2013","DOI":"10.1126\/scitranslmed.aaj2013","article-title":"Molecular remission of infant B-ALL after infusion of universal TALEN gene-edited CAR T cells","volume":"9","author":"Qasim","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"B29","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1111\/imr.12133","article-title":"T lymphocytes targeting native receptors","volume":"257","author":"Rooney","year":"2014","journal-title":"Immunol. Rev."},{"key":"B30","doi-asserted-by":"publisher","first-page":"737","DOI":"10.1038\/s41598-017-00462-8","article-title":"CRISPR\/Cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells","volume":"7","author":"Rupp","year":"2017","journal-title":"Sci. Rep."},{"key":"B31","doi-asserted-by":"publisher","first-page":"5099","DOI":"10.7150\/thno.84946","article-title":"A dendritic\/tumor fusion cell vaccine enhances efficacy of nanobody-based CAR-T cells against solid tumor","volume":"13","author":"Sun","year":"2023","journal-title":"Theranostics"},{"key":"B32","doi-asserted-by":"publisher","first-page":"3499","DOI":"10.1158\/1078-0432.ccr-16-2138","article-title":"Vaccination targeting native receptors to enhance the function and proliferation of chimeric antigen receptor (CAR)-Modified T cells","volume":"23","author":"Tanaka","year":"2017","journal-title":"Clin. Cancer Res."},{"key":"B33","doi-asserted-by":"publisher","first-page":"2033528","DOI":"10.1080\/2162402x.2022.2033528","article-title":"Comparing CAR and TCR engineered T cell performance as a function of tumor cell exposure","volume":"11","author":"Wachsmann","year":"2022","journal-title":"OncoImmunology"},{"key":"B34","doi-asserted-by":"publisher","first-page":"2993","DOI":"10.1158\/1078-0432.ccr-14-2920","article-title":"CMVpp65 vaccine enhances the antitumor efficacy of adoptively transferred CD19-Redirected CMV-Specific T cells","volume":"21","author":"Wang","year":"2015","journal-title":"Clin. Cancer Res."},{"key":"B35","doi-asserted-by":"publisher","first-page":"782","DOI":"10.1001\/jama.2020.12839","article-title":"Pathophysiology, transmission, diagnosis, and treatment of coronavirus disease 2019 (COVID-19): a review","volume":"324","author":"Wiersinga","year":"2020","journal-title":"JAMA"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.clml.2025.11.009","article-title":"Persistent COVID-19 in patients with hematological malignancies: a focused review in the Omicron era","author":"Yasuda","year":"2025","journal-title":"Clin. Lymphoma Myeloma Leukemia"},{"key":"B37","doi-asserted-by":"publisher","first-page":"2377","DOI":"10.3390\/v14112377","article-title":"Patients with hematological malignancies treated with T-Cell or B-Cell immunotherapy remain at high risk of severe forms of COVID-19 in the Omicron era","volume":"14","author":"Zerbit","year":"2022","journal-title":"Viruses"}],"container-title":["Frontiers in Cell and Developmental Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2026.1772146\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T06:56:26Z","timestamp":1772088986000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2026.1772146\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,26]]},"references-count":37,"alternative-id":["10.3389\/fcell.2026.1772146"],"URL":"https:\/\/doi.org\/10.3389\/fcell.2026.1772146","relation":{},"ISSN":["2296-634X"],"issn-type":[{"value":"2296-634X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,26]]},"article-number":"1772146"}}