CD19-CARCD7− T cells maintain a predominantly CD4+ phenotype and have comparable antitumor activity to CD19-CARBulk T cells. (A) CD19-CAR expression on bulk T cells and CD7− T cells (N = 10, 68.97 ± 12.20% vs 73.75 ± 8.27%, P < .01 compared with NT). (B) Immunophenotype of CD19-CAR–transduced T cells on day 7 posttransduction as determined by flow cytometry (CD4 vs CD8, effector memory (EM): CCR7−, CD45RA−; central memory (CM): CCR7+, CD45RA−; naïve-like: CCR7+CD45RA+; EM T cells reexpress CD45RA, EMRA: CCR7−, CD45RA+) (N = 3-6, 2-way ANOVA, values between bulk T cells and CD7− T cells were significant, P < .01, CD4+ and CD8+ EM bulk vs CD7− T cells P < .05, CD8+ naïve-like NTBulk vs NTCD7−P < .01, NTBulk vs CD19CD7−P < .05, NTCD7− vs CD19Bulk−P < .05). (C-D) Luciferase-based cytotoxicity assay to determine antitumor activity. (C) CD19-CAR T cells against BV173 (CD19+) for 24 hours, T-cell controls: NTBulk and NTCD7− T cells (N = 4, ns between CD19-CARBulk or CD7−, P < .05 at all ratios compared with NT T cells, 2-way ANOVA). (D) CD19-CARCD7− T cells were cocultured with BV173 (CD19+) or CCRF cells (CD19−) (N = 4, P < .05 at all ratios, 2-way ANOVA). ns, not significant.
