Fig. 2: Targeting TCRs to Trac sustains engineered T cell function and obviates Treg expansion.
From: Germline T cell receptor exchange results in physiological T cell development and function
a Schematic of retroviral transduction (RV) of P14 T cells with Msln-specific TCRs. b Schematic of CRISPR/Cas9 + rAAV TCR knockin (KI) of polyclonal T cells with Msln-specific TCRs. c Donor TCR expression by CD4 T cells 5 days post activation. d Donor TCR expression by CD8 T cells 5 days post activation. e Quantification of c and d. n = 2 biologically independent experiments. f Frequency of Ki67+ Vβ9+ CD4 or CD8 T cells 5 days post activation. n = 2 biologically independent experiments. g Frequency of Foxp3+ Vβ9+ CD4 or CD8 T cells 5 days post activation. n = 2 biologically independent experiments. h Representative plots gated on live CD4+ Vβ9+ T cells. i Representative plots gated on live CD8+ Vβ9+ T cells. j Representative plots gated on CD4+ Vβ9+ T cells. Intracellular cytokine staining was assessed after the second (Stim 2) or third (Stim 3) restimulation in vitro with Msln peptide-pulsed irradiated syngeneic splenocytes and IL-2 + IL-7. k Representative plots gated on CD8+ Vβ9+ T cells. Intracellular cytokine staining was assessed as in j. Data are mean ± SEM in e–g. n = 3–6 mice per group. *p < 0.05, **p < 0.005, and ***p < 0.0005. One-way ANOVA with a Tukey’s post test.
