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Stock, S.* ; Fertig, L.* ; Menkhoff, V.D.* ; Strzalkowski, T.* ; Caruso, M.* ; Kobold, S.

Retrovirus-based manufacturing of chimeric antigen receptor-modified T cells for cancer therapy research.

In:. 525 B Street, Suite 1900, San Diego, Ca 92101-4495 Usa: Elsevier, 2024. 329-352 (Methods Cell Biol. ; 191)

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Treatment with autologous chimeric antigen receptor (CAR)-modified T cells can achieve outstanding clinical response rates in heavily pretreated patients with B and plasma cell malignancies. However, relapses occur, and they limit the efficacy of this promising treatment approach. The complex GMP-compliant production and high treatment costs cause that CAR T cells cannot yet be used in a broad population. Among others, CAR T cell therapy has evolved regarding vector design and manufacturing process. Optimal production of CAR T cells is not yet defined, far from being standardized. Quality, cellular composition and immunophenotype of the administered CAR T cells are influenced by the manufacturing protocol and therefore play a crucial role for therapeutic success. For the gene transfer, viral and non-viral strategies are available. Retrovirus-based protocols for CAR T cell production offer advantages in terms of stable gene integration, sufficient transduction efficiency, proven clinical success, and scalability. Here, we detail a retrovirus-based generation protocol of human CAR-modified T cells for experimental immunotherapeutic treatment of cancer cells. For the CAR generation, HEK-293-based packaging cell lines, CD3+ selection, CD3/CD28-coated bead-based activation and IL-2/IL-15-mediated expansion were used. This protocol can be applied for every possible CAR construct after being successfully transfected in HEK-293-based packaging cell lines.

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Publication type Article: Edited volume or book chapter

Keywords Adoptive T Cell Therapy ; Car T Cells ; Chimeric Antigen Receptor ; Immunotherapy ; Retrovirus; Vectors; Immunotherapy; Suspension; Il-2

ISSN (print) / ISBN 0091-679X

Journal Methods in Cell Biology

Quellenangaben Volume: 191, Pages: 329-352

Publisher Elsevier

Publishing Place 525 B Street, Suite 1900, San Diego, Ca 92101-4495 Usa

Reviewing status Peer reviewed

Institute(s) Unit for Clinical Pharmacology (KKG-EKLiP)

Grants Bruno and Helene Joster Foundation
Ernst-Jung-Stiftung
German Cancer Aid (AvantCAR.de)
Melanoma Research Alliance Grants
International Doctoral Program i-Target: Immunotargeting of Cancer - Elite Network of Bavaria
Hector Foundation
Marie Sklodowska-Curie Program Training Network for Optimizing Adoptive T Cell Therapy of Cancer - H2020 Program of the European Union
BioVec Pharma
Quebec Network for Cell, Tissue and Gene Therapy (TheCell)
Forderprogramm fur Forschung und Lehre (FoFoLe) of the Medical Faculty of the LMU Munich
Novartis InCa Forderpreis 2022 for young researchers
DKTK School of Oncology
LMU Munich's Institutional Strategy LMUexcellent within the framework of the German Excellence Initiative
Bundesministerium fur Bildung und Forschung
Deutsche Jose Carreras Leukamie-Stiftung
Bavarian Research Foundation (BAYCELLator)
Fritz Bender Foundation
Deutsche Forschungsgemeinschaft (DFG)
European Research Council
Wilhelm Sander-Stiftung
Bavarian Ministry for Economical Affairs
Go-Bio Initiative
Else Kroner-Fresenius-Stiftung