Fig. 2: CAR design and human and murine T Cell transduction.

a Schematic representation of the retroviral vector expressing the p32 CAR (F = FLAG, L = Linker, LTR = long terminal repeat) and its transduction efficiency in mouse b and human c T cells. The transduction efficiency was measured by mcherry positive cells and by FLAG cell surface expression. Untransduced T cells were used as control (mUT = mouse and hUT = human). Histograms are representative of three independent experiments. d, e Frequency of CD4+/ CD8+ T cells 3 days post transduction showing no significant (ns) variation after CAR transduction of mouse (d) and human (e) T cells. Graphs on the right panel represent the quantification of N = 4 mice and N = 4 human donors, respectively. f, g Activation marker expression 2 days after transduction of mouse (f) and human (g) lymphocytes. Graphs on the right panel represent the quantification of N = 4 mice and N = 4 human donors, respectively. h, i Quantification of representative exhaustion marker expression 4 days (h, mouse, N = 3) and 10 days (i, human, N = 3 donors) after initial activation. j Phenotypic analysis of human UT (hUT) and CAR+(hp32) T cells at 10 days post transduction showing the frequency of central memory T cells (Tcm, CD45RA−CCR7+), stem central memory T cells (Tscm, CD45RA + CCR7+), effector T cells (Teff, CD45RA + CCR7−) and effector memory T cells (Tem, CD45RA−CCR7−) in CD8+ and CD4+ T cells; N = 4 donors. Each dot in the graphs represents a donor’s average of three independent experiments and final data is presented as mean ± SEM. Statistical significance was determined using an unpaired, two-sided t test when comparing between two groups (d, e, g, i, j), and multiple comparisons ANOVA test when comparing more than two groups (f, h). Source data are provided as a Source Data file.