A new approach to CAR T-cell gene engineering and cultivation using piggyBac transposon in the presence of IL-4, IL-7 and IL-21
Jazyk angličtina Země Velká Británie, Anglie Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
29475789
DOI
10.1016/j.jcyt.2017.10.001
PII: S1465-3249(17)30713-2
Knihovny.cz E-zdroje
- Klíčová slova
- CAR T cells, IL-21, IL-4, IL-7, cancer immunotherapy, piggyBac transposon,
- MeSH
- aktivace lymfocytů účinky léků genetika MeSH
- buněčné kultury metody MeSH
- buňky PC-3 MeSH
- chimerické antigenní receptory genetika metabolismus MeSH
- elektroporace MeSH
- genetické vektory MeSH
- HEK293 buňky MeSH
- imunoterapie adoptivní metody MeSH
- interleukin-21 MeSH
- interleukin-4 farmakologie MeSH
- interleukin-7 farmakologie MeSH
- interleukiny farmakologie MeSH
- kultivované buňky MeSH
- Lentivirus genetika MeSH
- lidé MeSH
- proteinové inženýrství metody MeSH
- protinádorové vakcíny genetika imunologie MeSH
- receptory antigenů T-buněk genetika metabolismus MeSH
- T-lymfocyty * cytologie účinky léků imunologie metabolismus MeSH
- transdukce genetická metody MeSH
- transpozibilní elementy DNA genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- chimerické antigenní receptory MeSH
- interleukin-21 MeSH
- interleukin-4 MeSH
- interleukin-7 MeSH
- interleukiny MeSH
- protinádorové vakcíny MeSH
- receptory antigenů T-buněk MeSH
- transpozibilní elementy DNA MeSH
BACKGROUND AIMS: Clinical-grade chimeric antigenic receptor (CAR)19 T cells are routinely manufactured by lentiviral/retroviral (LV/RV) transduction of an anti-CD3/CD28 activated T cells, which are then propagated in a culture medium supplemented with interleukin (IL)-2. The use of LV/RVs for T-cell modification represents a manufacturing challenge due to the complexity of the transduction approach and the necessity of thorough quality control. METHODS: We present here a significantly improved protocol for CAR19 T-cell manufacture that is based on the electroporation of peripheral blood mononuclear cells with plasmid DNA encoding the piggyBac transposon/transposase vectors and their cultivation in the presence of cytokines IL-4, IL-7 and IL-21. RESULTS: We found that activation of the CAR receptor by either its cognate ligand (i.e., CD19 expressed on the surface of B cells) or anti-CAR antibody, followed by cultivation in the presence of cytokines IL-4 and IL-7, enables strong and highly selective expansion of functional CAR19 T cells, resulting in >90% CAR+ T cells. Addition of cytokine IL-21 to the mixture of IL-4 and IL-7 supported development of immature CAR19 T cells with central memory and stem cell memory phenotypes and expressing very low amounts of inhibitory receptors PD-1, LAG-3 and TIM-3. CONCLUSIONS: Our protocol provides a simple and cost-effective method for engineering high-quality T cells for adoptive therapies.
Institute of Hematology and Blood Transfusion Prague Czech Republic
Institute of Molecular Genetics Czech Academy of Sciences Prague Czech Republic
Citace poskytuje Crossref.org
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