Extended Data Fig. 6: T3 cells proliferate and control NALM6 ALL in vivo.
From: T cells targeted to TdT kill leukemic lymphoblasts while sparing normal lymphocytes
(a) BLI analysis of NSG mice on day 9 after NALM-6ffluc-eGFP cell injection, one day prior to T-cell therapy. Data were pooled from two independent experiments, untreated n = 11, 1G4 n = 9, T3 n = 11. Data analysis by one-way ANOVA with Tukey’s multiple comparison test. (b,c) The percentage of human CD3+ (b) and CD8+ cells (c) in peripheral blood (PB) of mice analyzed at indicated time points out of total CD45+ human and CD45+ mouse leukocytes. (d) Percent cells expressing T3 and 1G4 TCRs among human CD8+ T cells throughout the experiment. Data in b-d are shown as mean ± s.e.m., n = 5 (1G4) and n = 6 (T3). (e) Flow cytometry plots showing bone-marrow tumor burden in one untreated and one 1G4-treated mice at time of sacrifice (day 14), and in five T3-treated mice sacrificed at end of experiment (day 57). (f) Expression of HLA-A2 and TdT in leukemia cells detected in bone marrow of untreated and 1G4-treated mice at the time of sacrifice. Light gray histograms represent negative control. (g) BLI images of mice in T3-treated group on day 57. Two of the T3-treated mice died for unknown reasons on day 35 and day 55 after T cell therapy, following negative BLI and PB flow cytometry analysis 5 days earlier, respectively, indicating that death was not related to leukemia. (h) IFN-γ levels in sera from untreated (n = 5), 1G4 (n = 4) or T3 (n = 5) cell-treated mice on day 2 after T cell therapy, measured as mean fluorescence intensity (MFI) of IFN-γ capturing fluorescent beads. Statistical analysis by one way ANOVA with Tukey’s multiple comparison test. Box plots in (a) and (h) show interquartile range (25th to 75th percentile) with central bar indicating the median and whiskers indicating the range, while dots represent data from individual mice.
