1. Long-term stability of T-cell activation and transduction components critical to the processing of clinical batches of gene-engineered T cells.
- Author
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Lamers CH, van Elzakker P, van Steenbergen SC, Luider BA, Groot C, van Krimpen BA, Vulto A, Sleijfer S, Debets R, and Gratama JW
- Subjects
- Adult, Antigens, Neoplasm genetics, Antigens, Neoplasm immunology, Carbonic Anhydrase IX, Carbonic Anhydrases genetics, Carbonic Anhydrases immunology, Carcinoma, Renal Cell genetics, Carcinoma, Renal Cell immunology, Cell Engineering, Cell Line, Fibronectins administration & dosage, Humans, Kidney Neoplasms genetics, Kidney Neoplasms immunology, Male, Muromonab-CD3 administration & dosage, Recombinant Proteins administration & dosage, T-Lymphocytes cytology, T-Lymphocytes drug effects, Tumor Necrosis Factor Receptor Superfamily, Member 7 drug effects, Carcinoma, Renal Cell therapy, Cell- and Tissue-Based Therapy, Kidney Neoplasms therapy, Receptors, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell immunology, T-Lymphocytes immunology
- Abstract
Background Aims: The generation of gene-modified T cells for clinical adoptive T-cell therapy is challenged by the potential instability and concomitant high financial costs of critical T-cell activation and transduction components. As part of a clinical trial to treat patients with metastatic renal cell cancer with autologous T cells engineered with a chimeric antigen receptor (CAR) recognizing carboxy-anhydrase-IX (CAIX), we evaluated functional stability of the retroviral vector, T-cell activation agent Orthoclone OKT3 (Janssen-Cilag, Beerse, Belgium) monoclonal antibody (mAb) and the transduction promoting agent RetroNectin (Takara, Otsu, Japan)., Methods: Carboxy-anhydrase-IX chimeric antigen receptor retrovirus-containing culture supernatants (RTVsups) were generated from two packaging cell lines, Phoenix-Ampho (BioReliance, Sterling, UK) and PG13, and stored at -80°C over 10 years and 14 years. For Orthoclone OKT3 and RetroNectin, aliquots for single use were prepared and stored at -80°C. Transduction efficiencies of both batches of RTVsups were analyzed using the same lots of cryopreserved donor peripheral blood mononuclear cells, Orthoclone OKT3 and RetroNectin over time., Results: We revisit here an earlier report on the long-term functional stability of the RTVsup, observed to be 9 years, and demonstrate that this stability is at least 14 years. Also, we now demonstrate that Orthoclone OKT3 and RetroNectin are functionally stable for periods of at least 6 years and 10 years., Conclusions: High-cost critical components for adoptive T-cell therapy can be preserved for ≥10 years when prepared in aliquots for single use and stored at -80°C. These findings may significantly facilitate, and decrease the financial risks of, clinical application of gene-modified T cells in multicenter studies., (Copyright © 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2013
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