Clinical grade manufacturing of genetically modified, CAR-expressing NK-92 cells for the treatment of ErbB2-positive malignancies
| Autor: | Annette Romanski, Congcong Zhang, Erhard Seifried, Paulina Nowakowska, Halvard Bonig, Marcus Odendahl, Winfried S. Wels, Nicole Miller, Torsten Tonn |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Blood Platelets Cytotoxicity Immunologic Cancer Research Receptor ErbB-2 medicine.medical_treatment Immunology Clone (cell biology) Cell Culture Techniques Pharmacology Immunotherapy Adoptive Radiation Tolerance GZMB 03 medical and health sciences 0302 clinical medicine NK-92 Cell Line Tumor Neoplasms medicine Immunology and Allergy Potency Humans Cell Proliferation Clinical Trials as Topic business.industry Genetic Therapy Culture Media Killer Cells Natural 030104 developmental biology Cytokine Oncology Cell culture 030220 oncology & carcinogenesis Cytokines Platelet lysate business Cell bank |
| Zdroj: | Cancer immunology, immunotherapy : CII. 67(1) |
| ISSN: | 1432-0851 |
| Popis: | The NK-92/5.28.z cell line (also referred to as HER2.taNK) represents a stable, lentiviral-transduced clone of ErbB2 (HER2)-specific, second-generation CAR-expressing derivative of clinically applicable NK-92 cells. This study addresses manufacturing-related issues and aimed to develop a GMP-compliant protocol for the generation of NK-92/5.28.z therapeutic doses starting from a well-characterized GMP-compliant master cell bank. Commercially available GMP-grade culture media and supplements (fresh frozen plasma, platelet lysate) were evaluated for their ability to support expansion of NK-92/5.28.z. Irradiation sensitivity and cytokine release were also investigated. NK-92/5.28.z cells can be grown to clinically applicable cell doses of 5 × 108 cells/L in a 5-day batch culture without loss of viability and potency. X-Vivo 10 containing recombinant transferrin supplemented with 5% FFP and 500 IU/mL IL-2 in VueLife 750-C1 bags showed the best results. Platelet lysate was less suited to support NK-92/5.28.z proliferation. Irradiation with 10 Gy completely abrogated NK-92/5.28.z proliferation and preserved viability and potency for at least 24 h. NK-92/5.28.z showed higher baseline cytokine release compared to NK-92, which was significantly increased upon encountering ErbB2(+) targets [GZMB (twofold), IFN-γ (fourfold), IL-8 (24-fold) and IL-10 (fivefold)]. IL-6 was not released by NK cells, but was observed in some stimulated targets. Irradiation resulted in upregulation of IL-8 and downregulation of sFasL, while other cytokines were not impacted. Our concept suggests NK-92/5.28.z maintenance culture from which therapeutic doses up to 5 × 109 cells can be expanded in 10 L within 5 days. This established process is feasible to analyze NK-92/5.28.z in phase I/II trials. |
| Databáze: | OpenAIRE |
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