Anti-CD19 CARs eradicated lymphoma cells and normal B cells. (A) The 38c13 lymphoma expressed κ light chain but not B220. In contrast, normal splenocytes did not have a substantial population of cells that expressed κ light chain but not B220. Almost all normal B cells expressed both κ light chain and B220. The numbers on the plots are the percentages of live cells in each quadrant. (B) The 38c13 lymphoma expressed CD19 but not B220. Normal splenocytes did not have a substantial population of CD19-expressing B220⁻ cells. Normal B cells are CD19⁺B220⁺. The numbers on the plots are the percentages of live cells in each quadrant. (C) Mice received 5 Gy of TBI, and later the same day the mice were injected intraperitoneally with 38c13 cells. One day after the lymphoma injection, groups of mice were injected intravenously with either 1D3-28Z.1-3-transduced T cells or 1D3-28Z–transduced T cells. Each mouse was injected with 6 × 10⁶ total T cells. The mice received IL-2 once daily on the day of T-cell transfer and the next day. A control group that did not receive any T cells was also included. Eight days later, the mice were killed and splenocytes were analyzed by flow cytometry. The plots are gated on live lymphocytes. The numbers on the plots are the percentages of live cells in each quadrant. Nearly identical results were obtained in 2 separate experiments in which 5 mice were injected with either 1D3-28Z.1-3–transduced T cells or 1D3-28Z–transduced T cells. (D) Mice received 5 Gy of TBI, and later the same day the mice were injected intraperitoneally with 38c13 cells. The next day, groups of mice were injected intravenously with T cells that were transduced with either the anti-CD19 CAR 1D3-28Z.1-3 or the negative-control CAR SP6-28Z.1-3. The mice received IL-2 once daily on the day of T-cell transfer and the next day. The mice were killed 8 days after T-cell transfer. Splenocytes were stained with anti-κ light chain and B220 to detect normal B cells. The graph summarizes the numbers of splenic B cells from mice in a single experiment in which 3 mice received 1D3-28Z.1-3–transduced T cells and 4 mice received SP6-28Z.1-3–transduced T cells. Nearly identical results were obtained in 2 separate experiments. (E) Mice were irradiated and then injected intraperitoneally with 38c13 lymphoma. The next day, the mice received 1D3-28Z.1-3–transduced T cells. The mice received IL-2 on the day of T-cell transfer and the next day. One hundred forty-three days later, the mice were killed and splenocytes were analyzed for B cells and T cells by flow cytometry. A representative example of 5 mice tested is shown. The CD19 versus B220 plot and the κ light chain versus B220 plot are gated on live lymphocytes. The CD8 versus CD4 plot is gated on CD3⁺ cells. The numbers on the plots are the percentages of cells in each quadrant.