Abstract
Anti-CD19 Chimeric Antigen Receptor (CAR-) T cell therapies have shown
promise for treating B cell malignancies, but the clinical outcome is
influenced by both the CAR-T product and the patient’s immune system.
The role of Tγδ cells in the context of CAR-T cell therapy remains
poorly understood. This study investigates the clonal expansion,
transcriptional heterogeneity, and migration profiles of Tgd cells in
patients undergoing anti-CD19 CAR-T cell therapy. Longitudinal single
cell multi-omics analysis was performed on Tgd cells from four patients
receiving anti-CD19 CAR-T cell therapy. Single cell RNA-seq,
antibody-based protein profiling (AbSeq), and full-length TCRγδ
sequences revealed clonally expanded populations displaying plasticity
in T cell differentiation, and temporal dynamics of large clones,
suggesting ongoing expansion and differentiation. Clonally expanded Tγδ
cells had heterogeneous gene expression profiles, occupying seven
transcriptionally distinct clusters. Analysis of chemokine markers
indicated cluster-specific homing tendencies of circulating Tγδ cells to
peripheral tissues. We found unexpectedly high frequencies of Vδ1 and
Vδ3 cells in the blood with distinct gene and protein expression
profiles. This analysis provides insights into the dynamic and
heterogeneous nature of Tγδ cells following anti-CD19 CAR-T cell
therapy, contributing valuable information for optimizing CAR-T cell
therapies in B cell malignancies.