0011906 2019 RIV GB eng J
Ptáčková, Pavlína - Musil, Jan - Štach, Martin - Lesný, Petr - Němečková, Šárka - Král, V. - Fábry, M. - Otáhal, Pavel^G
A new approach to CAR T cell gene engineering using piggyBac transposon and cultivation in the presence of IL-4, IL-7 and IL-21.
Cytotherapy. Vol. 20, no. 4 (2018), s. 507-520. ISSN 1465-3249
Grant: Ministerstvo zdravotnictví NV15-34498A; Ministerstvo zdravotnictví IP
Klíčová slova: CAR T cells * cancer immunotherapy * IL-4 * IL-7 * IL-21 * piggyBac transposon
Cite Score: 7.000, rok: 2018; SJR: 1.287, rok: 2018; IF: 4.297, rok: 2018
DOI: 10.1016/j.jcyt.2017.10.001

Anotace: Clinical-grade CAR19 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 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 a thorough quality control.
We present here a significantly improved protocol for CAR19 T cell manufacture which is based on the electroporation of PBMCs with plasmid DNA encoding the piggyBac transposon/transposase vectors and their cultivation in the presence of cytokines IL-4, IL-7 and IL-21. We found that activation of CAR receptor either by its cognate ligand (i.e. CD19 expressed on the surface of B cells) or by 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 more than 90% CAR+T cells.