Fig. 5. Vβ5-CAR-T cells were effective in vitro and in vivo against T cell leukemia.

a The expression levels of the TCR Vβ5 subtypes in untransduced T cells or Vβ5-specific CAR-T cells were analyzed by flow cytometry. b, c Untransduced T cells or Vβ5-CAR-T cells were cocultured with CCRF-CEM-Vβ5⁺ or Hut-78-Vβ5⁺ cells in triplicate wells at a CAR-T:tumor cell ratio of 2:1 for 24 h (n = 3 biologically independent samples/group). The remaining Vβ5⁺ cells, was determined by flow cytometry (b), and the indicated cytokines in the supernatants were analyzed by performing CBA assays (c). d female NSG mice aged 6–8 weeks (n = 7 mice/group) were intravenously inoculated with 1 × 10⁶ CCRF-CEM-Vβ5⁺ cells. Seven days later, tumor-bearing mice were treated with 1 × 10⁷ untransduced T cells or Vβ5-CAR-T cells. Nine and nineteen days after treatment, PBMCs were collected to analyze the CCRF-CEM-Vβ5⁺ (hCD45⁺Vβ5⁺) tumor cell burden. Statistical significance was determined by two-sided unpaired t tests. The normality of the data was confirmed by the Shapiro–Wilk test. Representative results of one from two replicate experiments are shown. The data shown represent the mean ± SEM (b-d). e Kaplan–Meier OS curves are shown for the inoculated mice (n = 9 mice for control T group and n = 6 mice for Vβ5-CAR-T group). Statistical significance was determined by the two-sided Mantel–Cox test. Representative results of one from two replicate experiments are shown.