CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells

Area of Science:

• Immunology
• Cell Therapy
• Biotechnology

Background:

• The Reverse Chimeric Antigen Receptor (RevCAR) system offers tunable T cell activity for improved safety in adoptive cell therapy.
• On-target, off-tumor toxicities remain a challenge in CAR T cell therapies.
• Dual-RevCAR T cells, utilizing two RevCAR constructs, enable AND-gate targeting for enhanced specificity.

Purpose of the Study:

• To evaluate the automated, clinical-grade manufacturing of RevCAR and Dual-RevCAR T cells using the CliniMACS Prodigy.
• To assess the expansion, phenotype, and functionality of RevCAR and Dual-RevCAR T cells produced via automated processing.
• To demonstrate the RevTM-dependent activity of these engineered T cells for potential clinical applications.

Main Methods:

• Automated processing of RevCAR and Dual-RevCAR T cells using the CliniMACS Prodigy system.
• Assessment of cell expansion, transduction efficiency, and T cell phenotype.
• Functional assays to evaluate RevTM-dependent cytotoxicity in both monospecific and AND-gate targeting strategies.

Main Results:

• Automated manufacturing via CliniMACS Prodigy yielded potent expansion and strong transduction of RevCAR and Dual-RevCAR T cells.
• The produced cell products exhibited a favorable phenotype suitable for clinical application.
• Functional studies confirmed strict RevTM-dependent activity for both monospecific and AND-gate targeting approaches.

Conclusions:

• Automated processing of RevCAR and Dual-RevCAR T cells is feasible and yields high-quality cell products.
• The RevCAR system, particularly Dual-RevCAR T cells, demonstrates significant potential for precise and safe tumor targeting in clinical settings.
• This automated manufacturing approach supports the clinical translation of advanced T cell therapies.