Enriching leukapheresis improves T cell activation and transduction efficiency during CAR T processing
Elsa Noaks1, Carlotta Peticone2, Ekaterini Kotsopoulou2
1Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK.
Molecular Therapy. Methods & Clinical Development
|March 15, 2021
Summary
Removing monocytes from leukapheresis material significantly enhances chimeric antigen receptor (CAR) T-cell production. This optimization improves CAR T-cell activation, transduction efficiency, and cytotoxic function, leading to a more potent and consistent therapeutic product.
Area of Science:
- Cellular immunotherapy
- CAR T-cell manufacturing
Background:
- Autologous manufacturing is standard for CD19-directed CAR T-cell products.
- Patient leukapheresis introduces variability and inhibitory cell populations, like myeloid cells, impacting CAR T-cell production.
Purpose of the Study:
- To investigate the impact of monocyte (CD14) depletion on CAR T-cell activation, growth, and transduction efficiency.
- To compare CAR T-cell production using different platforms (well plate, culture bag) and starting materials.
Main Methods:
- Healthy donor leukapheresis was used to assess the effect of monocyte depletion.
- Monocytes were removed from starting material prior to CAR T-cell culture.
- Activation levels, transduction efficiency, gene expression, and cytotoxicity were analyzed.
Main Results:
- Monocyte removal improved T-cell activation twofold and enhanced transduction efficiency by up to 64% in culture bag systems.
- CAR T-cell products from monocyte-depleted material showed superior cytotoxic activity.
- Depleted starting material yielded CAR T-cells with a more naive and rested memory phenotype.
Conclusions:
- Monocyte depletion is a critical step for optimizing autologous CAR T-cell manufacturing.
- This strategy enhances CAR T-cell potency, consistency, and phenotype, potentially improving therapeutic outcomes.
