Fig. 5: SEED-Selection enables the enrichment of TCR-swapped, coreceptor-swapped cells.
From: SEED-Selection enables high-efficiency enrichment of primary T cells edited at multiple loci
a, Diagram of a CD4 intron-targeted SEED HDRT encoding CD8α/β. b,c, CD4+ T cells were edited with a CD8 SEED (a) (+M3814) and immunomagnetically purified with anti-CD4 (n = 3 donors). b, Flow cytometry plots of CD4 and CD8 expression. c, Percentage of CD4+ and CD4−CD8+ cells. d–h CD4+ T cells were edited with 1G4-LY and CD8 SEEDs (+M3814) and immunomagnetically purified with BW242 and anti-CD4 (n = 3 donors). d, Overview of editing and selection strategy. e, Flow cytometry plots of CD4 and CD8 expression and BW242 and NY-ESO-1 dextramer binding. f, Percentage of cells that express CD4 or an endogenous or mispaired TCR (BW242+). Percentage of fully edited cells with minimal mispairing (BW242−CD4−dextramer+CD8+). g, Assessment of NY-ESO-1 dextramer binding by flow cytometry in CD4+CD8−BW242−1G4-LY+ cells (red), CD4−CD8+BW242−1G4-LY+ cells (blue) and nonedited CD4+ T cells (gray) (n = 3 donors). Data are displayed as the mean ± s.e.m. h, Assessment of cytotoxic activity of CD4+ T cells against NY-ESO-1+ A375 target cells (technical triplicate; data shown from one of four donors). Nonedited T cells, dark gray; 1G4-LY SEED-edited, red; 1G4-LY SEED-edited; CD8 SEED-edited, blue. Data are displayed as the mean (bold line) ± s.e.m. (shaded area). Significance was assessed at 72 h. The significance in g,h was assessed using a repeated-measures one-way ANOVA and Turkey’s multiple-comparisons test.
