Fig. 6: T cell cytotoxicity of select PSA-specific clonotypes from ATLAS-seq.

A ATLAS-seq selected clonotypes sorted according to clonotype proportions (top) and cytotoxic cytokine indexes (bottom). PSA-specific TCRs used for target cell killing assay are highlighted and labeled. B Target cell killing assays: TCR-transduced PBMCs were activated and then cocultured for 72 h with GFP expressing HLA-A*02:01 cell lines LNCaP (PSA positive target) or PC3 cells (PSA negative control). Relative viability of target cells was measured by GFP fluorescence using the Incucyte system. Each dot: mean of n = 3 independent biological replicates. C Images of representative GFP/ HLA-A*02:01 -expressing LNCaP and PC3 cells after 72-h coculture with TCR P1 and P40 transduced PBMCs or un-transduced (UT) PBMCs. Target cells are shown in green. Scale bar: 400μm. D Secreted IFNγ and TNF concentration after the 72-h cell killing assay. Conditioned media in the same wells were measured by IFNγ and TNF ELISA (mean ± s.d. of n = 3 independent biological replicates). +PC3: The mean measurement of cytokines in the conditioned media of cocultures of PC3 cells (PSA negative control) with all 11 tested clonotypes. Source data are provided as a Source Data file.