In Vitro Expanded Cord Blood ?d T Cells Acquired Unique Cell State Composition to Mediate Sustained Human AML Suppression [VDJ]

Single cell TCR sequencing was performed to examine the clonal type changes in cord blood derived gamma-delta T-cells following in-vitro expansion in a co-culture with EBV-LCL feeder over 2 weeks. Following 2 weeks of expansion, the cells were challenged with the AML cell line K562. Single-cell TCR was performed after 24 hours post-challenge to identify clones that were preferentially expanded, as well as the corresponding cell states mapped using single-cell RNA-sequencing. Overall design: Cultured gamma-delta T-cells were sorted with FACS, cultured in the presence of in RPMI + 10% FBS + 170 U/mL recombinant human interleukin-2 (rhIL-2), 25U/ml rhIL-15 as well as 0.7U/ml rhIL-21, with EBV-LCL feeder over a period of 2 weeks and analyzed using scRNAseq before/after antigen challenge with K562.