Anti-CD19 (1D3) h(CD28-CD3ζ) CAR, pCDCAR1 (CAR-MZ020)

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.1 Flow cytometry analysis of CD3+ or CD19+ cells in mice treated with anti-CD19 (1D3) CAR-T or anti-FITC CAR-T with anti-CD19 (1D3)-FITC switch.

C57BL/6 mice were preconditioned with cyclophosphamide (150 mg/kg) on day 1, and received 6 × 106 syngeneic anti-CD19 (ID3) CAR or anti-FITC CAR-T cells by tail-vein injections the next day. Daily treatments with anti-mouse CD19 (1D3)-FITC switch at 1 mg/kg were initiated the same day as CAR-T-cell infusions for a total of 10 injections (days 2-11).

Ma, J. S., Kim, J. Y., Kazane, S. A., Choi, S. H., Yun, H. Y., Kim, M. S., ... & Cao, Y. (2016). Versatile strategy for controlling the specificity and activity of engineered T cells. Proceedings of the National Academy of Sciences, 113(4), E450-E458.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.2 Quantified analysis of CD19+ cells in mice treated with anti-CD19 (1D3) CAR-T or anti-FITC CAR-T with anti-CD19 (1D3)-FITC switch.

Graphical representation of CD19+ cells quantified from B. Results are displayed as an average of five to six mice.

Ma, J. S., Kim, J. Y., Kazane, S. A., Choi, S. H., Yun, H. Y., Kim, M. S., ... & Cao, Y. (2016). Versatile strategy for controlling the specificity and activity of engineered T cells. Proceedings of the National Academy of Sciences, 113(4), E450-E458.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.3 Anti-CD19–CAR-expressing T cells produce IL-2 and proliferate in response to CD19.

1D3-28Z.1-3-transduced T cells, 1D3-28Z-transduced T cells, or SP6-28Z.1-3-transduced T cells were cultured overnight alone or with the indicated target cells, and an IL-2 ELISA was performed.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.4 Functional anti-CD19–CAR-transduced T cells were detected in mice after adoptive transfer.

Mice received 5 Gy of TBI and injected intraperitoneally with 38c13 lymphoma cells, then received CAR-T cells . Eight days after the T-cell transfer, the mice were killed and splenocytes were stained with mouse anti-rat Fab antibodies to detect CAR-transduced cells.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.5 Functional anti-CD19–CAR-transduced T cells were detected in mice after adoptive transfer.

Mean absolute numbers of CAR-expressing CD3+CD8+ splenocytes in mice that received either 1D3-28Z.1-3-transduced T cells or 1D3-28Z-transduced T cells.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.6 Functional anti-CD19–CAR-transduced T cells were detected in mice after adoptive transfer.

Mean absolute numbers of CAR-expressing CD3+CD4+ splenocytes in mice that received either 1D3-28Z.1-3-transduced T cells or 1D3-28Z-transduced T cells.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.7 Functional anti-CD19–CAR-transduced T cells were detected in mice after adoptive transfer.

Eight days after T-cell transfer, the mice were killed, and splenocytes were cultured with 38c13, CD19-K562, or NGFR-K562. 38c13 and CD19-K562 were CD19+. NGFR-K562 was CD19-. Intracellular cytokine staining was performed for IFN-γ. The numbers on the plots represent the percentage of CD3+ cells.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.8 Anti-CD19–CAR-transduced T cells eradicated large lymphoma masses

Mice received 5 Gy of TBI and injected intraperitoneally with 38c13 lymphoma cells, then received CAR-T cells. The mean tumor sizes of each group are shown.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.

CAR Construction : 1D3 scfv-CD28-CD3ζ

Fig.9 Anti-CD19–CAR-transduced T cells eradicated large lymphoma masses

The survival of the same groups of mice described in vivo assay is shown.

Kochenderfer, J. N., Yu, Z., Frasheri, D., Restifo, N. P., & Rosenberg, S. A. (2010). Adoptive transfer of syngeneic T cells transduced with a chimeric antigen receptor that recognizes murine CD19 can eradicate lymphoma and normal B cells. Blood, The Journal of the American Society of Hematology, 116(19), 3875-3886.