CAR T cell engineering impacts antigen-independent activation and co-inhibition.

ABSTRACT: Viral vectors have successfully modified T cells to express chimeric antigen receptors (CARs), leading to clinical approvals. However, their high cost and regulatory challenges hinder rapid clinical translation. Here, we demonstrate that our lentivirally (LV) manufactured R110-CAR T cells, targeting a leukemia neoepitope, can also be engineered using the non-viral sleeping beauty (SB) transposition with minimal-sized DNA vectors. Flow cytometry and single-cell sequencing were used to compare the two production modes using healthy donor and chronic lymphocytic leukemia (CLL)-patient-derived T cells and a CD19-CAR T cell control. SB products shifted toward CD8⁺ subsets with activation/co-inhibition marker expression (CD69, LAG-3, and TIM-3) despite their naïve-like phenotype and l

SUBMITTER: Schultheiß C 

PROVIDER: S-EPMC12464704 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature