Fig. 2: AAV-CD4CAR induces antitumor activity.
From: AAV-mediated in vivo CAR gene therapy for targeting human T-cell leukemia
A Schematic representation showing the ex vivo AAV-CAR production and co-culture assays. B AAV-CD4CAR depleted CD3+CD4+ cells in different donors (n = 10) during the course of T-cell expansion. Activated PBMCs from each donor were transduced with different doses of AAV-CD4CAR, and Vector control (untransduced). After 48 h, the cells were analyzed by FACS using anti-human CD4, anti-human CD3, and anti-human CD8 antibodies. The CD3+CD4+ percentage was normalized to the vector control group (which represented 100%), and the CD3+CD4+ percentages of AAV-CD4CAR (H) and AAV-CD4CAR (L) was calculated by the percent difference from the vector control group. A two-way ANOVA multiple comparisons test within each donor was used for statistical analysis. ns, no significance; ***P < 0.001; **P < 0.01; *P < 0.05. C AAV-CD4CAR T cells were co-cultured with CFSE-stained target cells, 48 h post-co-culture, cells were stained with 7-AAD and analyzed by FACS. Two-way ANOVA with the multiple comparisons test was used to assess significance, ns, no significance; ****P < 0.0001. ***P < 0.001; **P < 0.01; *P < 0.05. D ATL patient PBMCs were infected with AAV-CD4CAR (H) and with a control vector (samples were duplicated). Forty-eight hours post-transduction, cells were analyzed by FACS for CD3+CD4+ depletion. An unpaired two-tailed t-test was used to calculate the significance of the difference. AAV-CD4CAR vs vector control, ***P < 0.0001.
