Our center performs experimental clinical studies with advanced therapy medicinal products (ATMPs) based on polyclonal T cells, all of which are currently expanded in standard T-flasks. Given the need to increase the efficiency and safety of large-scale T cell expansion for clinical use, we have optimized the method to expand in G-Rex devices both cytokine-induced killer cells (CIKs) from peripheral or cord blood and blinatumomab-expanded T cells (BETs). We show that the G-Rex reproducibly allowed the expansion of >30 × 106 CD3+ cells/cm2 of gas-permeable membrane in a mean of 10 to 11 days in a single unit, without manipulation, except for addition of cytokines and sampling of supernatant for lactate measurement every 3 to 4 days. In contrast, 21 to 24 days, twice-weekly cell resuspension and dilution into 48 to 72 T-flasks were required to complete expansions using the standard method. We show that the CIKs produced in G-Rex (CIK-G) were phenotypically very similar, for a large panel of markers, to those expanded in T-flasks, although CIK-G products had lower expression of CD56 and higher expression of CD27 and CD28. Functionally, CIK-Gs were strongly cytotoxic in vitro against the NK cell target K562 and the REH pre-B ALL cell line in the presence of blinatumomab. CIK-Gs also showed therapeutic activity in vivo in the Ph+ pre-B ALL-2 model in mice. The expansion of both CIKs and BETs in G-Rex was validated in good manufacturing practices (GMP) conditions, and we plan to use G-Rex for T cell expansion in future clinical studies.

Gotti, E., Tettamanti, S., Zaninelli, S., Cuofano, C., Cattaneo, I., Rotiroti, M., et al. (2022). Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use. CYTOTHERAPY, 24(3 (March 2022)), 334-343 [10.1016/j.jcyt.2021.11.004].

Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use

Tettamanti S.;Zaninelli S.;Cattaneo I.;Rotiroti M. C.;Rambaldi A.;Introna M.
;
2022

Abstract

Our center performs experimental clinical studies with advanced therapy medicinal products (ATMPs) based on polyclonal T cells, all of which are currently expanded in standard T-flasks. Given the need to increase the efficiency and safety of large-scale T cell expansion for clinical use, we have optimized the method to expand in G-Rex devices both cytokine-induced killer cells (CIKs) from peripheral or cord blood and blinatumomab-expanded T cells (BETs). We show that the G-Rex reproducibly allowed the expansion of >30 × 106 CD3+ cells/cm2 of gas-permeable membrane in a mean of 10 to 11 days in a single unit, without manipulation, except for addition of cytokines and sampling of supernatant for lactate measurement every 3 to 4 days. In contrast, 21 to 24 days, twice-weekly cell resuspension and dilution into 48 to 72 T-flasks were required to complete expansions using the standard method. We show that the CIKs produced in G-Rex (CIK-G) were phenotypically very similar, for a large panel of markers, to those expanded in T-flasks, although CIK-G products had lower expression of CD56 and higher expression of CD27 and CD28. Functionally, CIK-Gs were strongly cytotoxic in vitro against the NK cell target K562 and the REH pre-B ALL cell line in the presence of blinatumomab. CIK-Gs also showed therapeutic activity in vivo in the Ph+ pre-B ALL-2 model in mice. The expansion of both CIKs and BETs in G-Rex was validated in good manufacturing practices (GMP) conditions, and we plan to use G-Rex for T cell expansion in future clinical studies.
Articolo in rivista - Articolo scientifico
bioreactors; cytokine-induced killer cells; immunotherapy; T cells;
English
334
343
10
Gotti, E., Tettamanti, S., Zaninelli, S., Cuofano, C., Cattaneo, I., Rotiroti, M., et al. (2022). Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use. CYTOTHERAPY, 24(3 (March 2022)), 334-343 [10.1016/j.jcyt.2021.11.004].
Gotti, E; Tettamanti, S; Zaninelli, S; Cuofano, C; Cattaneo, I; Rotiroti, M; Cribioli, S; Alzani, R; Rambaldi, A; Introna, M; Golay, J
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/347944
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