{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T12:14:34Z","timestamp":1792671274041,"version":"4.5.0"},"reference-count":32,"publisher":"American Society of Hematology","issue":"12","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2020,9,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Anti-CD19 chimeric antigen receptor (CAR) T cells showed significant antileukemic activity in B-precursor acute lymphoblastic leukemia (ALL). Allogeneic, HLA-mismatched off-the-shelf third-party donors may offer ideal fitness of the effector cells, but carry the risk of graft-versus-host disease. Knockout (KO) of the endogenous T-cell receptor (TCR) in CD19-CAR-T cells may be a promising solution. Here, we induced a CRISPR\/Cas9-mediated KO of the TCR\u03b2 chain in combination with a second-generation retroviral CAR transduction including a 4-1BB costimulatory domain in primary T cells. This tandem engineering led to a highly functional population of TCR-KO-CAR-T cells with strong activation (CD25, interferon \u03b3), proliferation, and specific killing upon CD19 target recognition. TCR-KO-CAR-T cells had a balanced phenotype of central memory and effector memory T cells. KO of the endogenous TCR in T cells strongly ablated alloreactivity in comparison with TCR-expressing T cells. In a patient-derived xenograft model of childhood ALL, TCR-KO-CAR-T cells clearly controlled CD19+ leukemia burden and improved survival in vivo. However, coexpression of endogenous TCR plus CAR led to superior persistence of T cells and significantly prolonged leukemia control in vivo, confirmed by a second in vivo model using the leukemia cell line NALM6. These results point toward an essential role of the endogenous TCR for longevity of the response at the price of alloreactivity. In conclusion, anti-CD19 CAR T cells with a CRISPR\/Cas9-mediated TCR-KO are promising candidates for nonmatched third-party adoptive T-cell transfer with high antileukemic functionality in the absence of alloreactivity, but long-term persistence in vivo is better in the presence of the endogenous TCR.<\/jats:p>","DOI":"10.1182\/blood.2020005185","type":"journal-article","created":{"date-parts":[[2020,6,3]],"date-time":"2020-06-03T02:51:09Z","timestamp":1591152669000},"page":"1407-1418","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":181,"title":["Endogenous TCR promotes in vivo persistence of CD19-CAR-T cells compared to a CRISPR\/Cas9-mediated TCR knockout CAR"],"prefix":"10.1182","volume":"136","author":[{"given":"Dana","family":"Stenger","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"},{"name":"German Cancer Consortium, partner site, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanja A.","family":"Stief","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"},{"name":"National Center for Infection Research, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Theresa","family":"Kaeuferle","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9472-2191","authenticated-orcid":false,"given":"Semjon","family":"Willier","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felicitas","family":"Rataj","sequence":"additional","affiliation":[{"name":"Center for Integrated Protein Science Munich and Division of Clinical Pharmacology, Department of Medicine IV, Klinikum der Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9323-9472","authenticated-orcid":false,"given":"Kilian","family":"Schober","sequence":"additional","affiliation":[{"name":"National Center for Infection Research, Munich, Germany;"},{"name":"Institute for Medical Microbiology, Immunology and Hygiene, Technische Universit\u00e4t M\u00fcnchen, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1956-2778","authenticated-orcid":false,"given":"Binje","family":"Vick","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"},{"name":"German Cancer Consortium, partner site, Munich, Germany;"},{"name":"Research Unit Apoptosis in Hematopoietic Stem Cells, Helmholtz Zentrum M\u00fcnchen, German Research Center for Environmental Health, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8125-0591","authenticated-orcid":false,"given":"Ramin","family":"Lotfi","sequence":"additional","affiliation":[{"name":"Institute for Transfusion Medicine, University Hospital Ulm, Ulm, Germany;"},{"name":"German Red Cross Blood Services Baden-W\u00fcrttemberg-Hessen, Institute for Clinical Transfusion Medicine and Immunogenetics Ulm, Ulm, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beate","family":"Wagner","sequence":"additional","affiliation":[{"name":"Department for Transfusion Medicine, Cell Therapeutics and Hemostaseology, Klinikum der Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0920-7377","authenticated-orcid":false,"given":"Thomas G. P.","family":"Gr\u00fcnewald","sequence":"additional","affiliation":[{"name":"German Cancer Consortium, partner site, Munich, Germany;"},{"name":"Max-Eder Research Group for Pediatric Sarcoma Biology, Institute of Pathology, Ludwig Maximilians University Munich, Munich, Germany;"},{"name":"German Cancer Research Center, Heidelberg, Germany; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebastian","family":"Kobold","sequence":"additional","affiliation":[{"name":"Center for Integrated Protein Science Munich and Division of Clinical Pharmacology, Department of Medicine IV, Klinikum der Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dirk H.","family":"Busch","sequence":"additional","affiliation":[{"name":"National Center for Infection Research, Munich, Germany;"},{"name":"Institute for Medical Microbiology, Immunology and Hygiene, Technische Universit\u00e4t M\u00fcnchen, Munich, Germany;"},{"name":"Focus Group \u201cClinical Cell Processing and Purification,\u201d Institute for Advanced Study, Technische Universit\u00e4t M\u00fcnchen, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1773-7677","authenticated-orcid":false,"given":"Irmela","family":"Jeremias","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"},{"name":"German Cancer Consortium, partner site, Munich, Germany;"},{"name":"Research Unit Apoptosis in Hematopoietic Stem Cells, Helmholtz Zentrum M\u00fcnchen, German Research Center for Environmental Health, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5770-4744","authenticated-orcid":false,"given":"Franziska","family":"Blaeschke","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8517-9681","authenticated-orcid":false,"given":"Tobias","family":"Feuchtinger","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children\u2019s Hospital, University Hospital, Ludwig Maximilians University Munich, Munich, Germany;"},{"name":"German Cancer Consortium, partner site, Munich, Germany;"},{"name":"National Center for Infection Research, Munich, 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