Fig. 7

Residues L96α, R9β, and Y10β enhance antigen-specific effector functions. Human T cells transduced with wild-type or L96α, R9β, and Y10β (LRY)-modified TCRs were stimulated with peptide-loaded T2 cells. a A representative example of n = 3 independent experiments showing the frequencies of gated CD19^high T cells that produced IFNγ and/or IL2. b Pooled data (means ± SEM) showing the fold increase in total specific cytokine production by LRY-modified TCR-transduced cells over the corresponding wild-type TCR-transduced cells. n = 3 independent experiments. Cytokine produced by stimulation with the irrelevant peptide were subtracted from cytokine produced by cognate peptide. c Transduced T cells labeled with Cell Trace Violet were co-cultured for 5 days with peptide-loaded T2 cells. Shown are the percentages of wild-type or LRY-modified TCR-transduced CD8⁺ T cells that underwent antigen-specific proliferation. Proliferation arising from irrelevant peptide stimulation was subtracted from cognate peptide-induced proliferation. n = 3 independent experiments. d The indicated transduced T cells were co-cultured overnight with T2 cells pulsed with control peptide or cognate peptide. Shown is the antigen-specific killing of n = 7 independent experiments for the CMV1 TCR and n = 6 independent experiments for the HA1.m2 and HA2.19 TCRs. e Transduced T cells were co-cultured for 4 h with peptide-loaded T2 cells. Shown are n = 3 independent experiments measuring antigen-specific upregulation of CD69 on CD8⁺ T cells expressing wild-type or LRY-modified TCRs. CD69 expression in response to irrelevant peptide stimulation was subtracted from cognate peptide induced CD69 expression. f Transduced T cells were stimulated overnight with T2 cells loaded with the indicated concentrations of cognate peptide. IFNγ production was measured by ELISA. Data were pooled (means ± SEM) and normalized to IFNγ production by wild-type TCR-transduced T cells stimulated with T2 cells loaded with 10 μM cognate peptide. n = 3 independent experiments. g CMV1 TCR-transduced T cells were stimulated overnight with T2 cells expressing variant or cognate peptide. IFNγ production was measured by ELISA. Data were pooled (means ± SEM) and normalized to IFNγ produced in response to cognate peptide stimulation. n = 3 independent experiments. ns (non-significant), P > 0.05 (Mann–Whitney U test) for all comparisons between the LRY-modified TCR and the wild-type TCR