PMID- 34058386 OWN - NLM STAT- MEDLINE DCOM- 20220407 LR - 20230106 IS - 1525-0024 (Electronic) IS - 1525-0016 (Print) IS - 1525-0016 (Linking) VI - 30 IP - 1 DP - 2022 Jan 5 TI - Protective T cell receptor identification for orthotopic reprogramming of immunity in refractory virus infections. PG - 198-208 LID - S1525-0016(21)00303-8 [pii] LID - 10.1016/j.ymthe.2021.05.021 [doi] AB - Viral infections cause life-threatening disease in immunocompromised patients and especially following transplantation. T cell receptor (TCR) engineering redirects specificity and can bring significant progress to emerging adoptive T cell transfer (ACT) approaches. T cell epitopes are well described, although knowledge is limited on which TCRs mediate protective immunity. In this study, refractory adenovirus (AdV) infection after hematopoietic stem cell transplantation (HSCT) was treated with ACT of highly purified Hexon5-specific T cells using peptide major histocompatibility complex (pMHC)-Streptamers against the immunodominant human leukocyte antigen (HLA)-A *0101-restricted peptide LTDLGQNLLY. AdV was successfully controlled through this oligoclonal ACT. Novel protective TCRs were isolated ex vivo and preclinically engineered into the TCR locus of allogeneic third-party primary T cells by CRISPR-Cas9-mediated orthotopic TCR replacement. Both TCR knockout and targeted integration of the new TCR in one single engineering step led to physiological expression of the transgenic TCR. Reprogrammed TCR-edited T cells showed strong virus-specific functionality such as cytokine release, effector marker upregulation, and proliferation capacity, as well as cytotoxicity against LTDLGQNLLY-presenting and AdV-infected targets. In conclusion, ex vivo isolated TCRs with clinical proven protection through ACT could be redirected into T cells from naive third-party donors. This approach ensures that transgenic TCRs are protective with potential off-the-shelf use and widened applicability of ACT to various refractory emerging viral infections. CI - Published by Elsevier Inc. FAU - Stief, Tanja A AU - Stief TA AD - Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital, LMU Munich, Munich Germany; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany. FAU - Kaeuferle, Theresa AU - Kaeuferle T AD - Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital, LMU Munich, Munich Germany; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany. FAU - Muller, Thomas R AU - Muller TR AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen (TUM), Munich, Germany; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany. FAU - Doring, Michaela AU - Doring M AD - Department I - General Pediatrics, Hematology/Oncology, University Hospital Tubingen, Children's Hospital, Tubingen, Germany. FAU - Jablonowski, Lena M AU - Jablonowski LM AD - Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital, LMU Munich, Munich Germany. FAU - Schober, Kilian AU - Schober K AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen (TUM), Munich, Germany; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany. FAU - Feucht, Judith AU - Feucht J AD - Department I - General Pediatrics, Hematology/Oncology, University Hospital Tubingen, Children's Hospital, Tubingen, Germany; Center for Cell Engineering, Memorial Sloan Kettering Cancer Center, New York, NY, USA. FAU - Dennehy, Kevin M AU - Dennehy KM AD - German Center for Infection Research (DZIF), Partner Site Tubingen, Tubingen, Germany; Institute for Laboratory Medicine and Microbiology, University Hospital Augsburg, Germany. FAU - Willier, Semjon AU - Willier S AD - Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital, LMU Munich, Munich Germany. FAU - Blaeschke, Franziska AU - Blaeschke F AD - Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital, LMU Munich, Munich Germany; Department I - General Pediatrics, Hematology/Oncology, University Hospital Tubingen, Children's Hospital, Tubingen, Germany. FAU - Handgretinger, Rupert AU - Handgretinger R AD - Department I - General Pediatrics, Hematology/Oncology, University Hospital Tubingen, Children's Hospital, Tubingen, Germany. FAU - Lang, Peter AU - Lang P AD - Department I - General Pediatrics, Hematology/Oncology, University Hospital Tubingen, Children's Hospital, Tubingen, Germany. FAU - Busch, Dirk H AU - Busch DH AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen (TUM), Munich, Germany; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany. FAU - Feuchtinger, Tobias AU - Feuchtinger T AD - Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital, LMU Munich, Munich Germany; Department I - General Pediatrics, Hematology/Oncology, University Hospital Tubingen, Children's Hospital, Tubingen, Germany; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany. Electronic address: tobias.feuchtinger@med.uni-muenchen.de. LA - eng GR - P30 CA008748/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210529 PL - United States TA - Mol Ther JT - Molecular therapy : the journal of the American Society of Gene Therapy JID - 100890581 RN - 0 (Receptors, Antigen, T-Cell) SB - IM MH - Adoptive Transfer MH - Humans MH - Immunotherapy, Adoptive MH - *Receptors, Antigen, T-Cell/genetics MH - T-Lymphocytes MH - *Virus Diseases PMC - PMC8753271 OTO - NOTNLM OT - AdV-specific T cells OT - CRISPR/Cas9 OT - adoptive T-cell transfer OT - homology-directed repair OT - orthotopic TCR replacement COIS- Declaration of interests The authors declare no competing interests. EDAT- 2021/06/01 06:00 MHDA- 2022/04/08 06:00 PMCR- 2023/01/05 CRDT- 2021/05/31 20:14 PHST- 2021/01/03 00:00 [received] PHST- 2021/04/30 00:00 [revised] PHST- 2021/05/25 00:00 [accepted] PHST- 2021/06/01 06:00 [pubmed] PHST- 2022/04/08 06:00 [medline] PHST- 2021/05/31 20:14 [entrez] PHST- 2023/01/05 00:00 [pmc-release] AID - S1525-0016(21)00303-8 [pii] AID - 10.1016/j.ymthe.2021.05.021 [doi] PST - ppublish SO - Mol Ther. 2022 Jan 5;30(1):198-208. doi: 10.1016/j.ymthe.2021.05.021. Epub 2021 May 29.