pCDCAR1 PSMA h(BBζ) (CAR-ZP053)

CAR Construction : 3D8 scFv-28ζ

Fig.1 Detection of PSMA cell surface expression on PC3-PSMA cells by 3D8 mAb.

PC3 and PC3-PSMA cellswere analyzedby flowcytometry staining with 3D8 mAb followed by incubation with secondary PE-conjugated anti-mouse IgG antibody (solid line).

Ma, Q., Gomes, E. M., Lo, A. S. Y., & Junghans, R. P. (2014). Advanced generation anti‐prostate specific membrane antigen designer T Cells for prostate cancer immunotherapy. The prostate, 74(3), 286-296.

CAR Construction : 3D8 scFv-28ζ

Fig.2 Expression of anti-PSMA CARs in T cells.

Expression of IgTCR and IgCD28 TCR.

Ma, Q., Gomes, E. M., Lo, A. S. Y., & Junghans, R. P. (2014). Advanced generation anti‐prostate specific membrane antigen designer T Cells for prostate cancer immunotherapy. The prostate, 74(3), 286-296.

CAR Construction : 3D8 scFv-28ζ

Fig.3 Homodimer formation of IgTCR and IgCD28 TCR.

Positions of monomer and dimer of TCRz, IgTCR and IgCD28 TCR are indicated on the right. Molecular mass markers (kDa) are indicated on the left.

Ma, Q., Gomes, E. M., Lo, A. S. Y., & Junghans, R. P. (2014). Advanced generation anti‐prostate specific membrane antigen designer T Cells for prostate cancer immunotherapy. The prostate, 74(3), 286-296.

CAR Construction : 3D8 scFv-28ζ

Fig.4 Superior cytokine production by 2ndgen dTc.Non-transduced,IgTCR andIgCD28 TCR transduced T cells were incubated with control PC3 and PC3-PSMA tumor cells for 24 hr.

Supernatants were harvested and assayed for IL2 and IFNg by ELISA.

Ma, Q., Gomes, E. M., Lo, A. S. Y., & Junghans, R. P. (2014). Advanced generation anti‐prostate specific membrane antigen designer T Cells for prostate cancer immunotherapy. The prostate, 74(3), 286-296.

CAR Construction : 3D8 scFv-28ζ

Fig.5 Proliferation of dTc in response to PSMAþ tumor cells.

Total cell proliferation.Non-transduced control,IgTCR and IgCD28TCR transduced T cells with 45% transduction efficiency for both were stimulated with g-irradiated PC3 and PC3-PSMA tumor cells for 3 days,cultured for a further 6 days, and counted on days noted.

Ma, Q., Gomes, E. M., Lo, A. S. Y., & Junghans, R. P. (2014). Advanced generation anti‐prostate specific membrane antigen designer T Cells for prostate cancer immunotherapy. The prostate, 74(3), 286-296.

CAR Construction : 3D8 scFv-28ζ

Fig.6 GD2 sugar moiety and correlation of anti-GD2 mAb affinity with clinical efficacy.

Data are represented as mean±s.d.

Hassani, M., Hajari Taheri, F., Sharifzadeh, Z., Arashkia, A., Hadjati, J., van Weerden, W. M., ... & Abolhassani, M. (2019). Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer. Journal of Cellular Biochemistry, 120(6), 10787-10795.

CAR Construction : 3D8 scFv-28ζ

Fig.7 Schematic presentation of NBPII‐CAR and expression of NBPII‐CAR on electroporated T cells.

Tumor cells were stained with PE-conjugated anti-PSMA antibody or isotype control antibody and analyzed by flow cytometry.

Hassani, M., Hajari Taheri, F., Sharifzadeh, Z., Arashkia, A., Hadjati, J., van Weerden, W. M., ... & Abolhassani, M. (2019). Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer. Journal of Cellular Biochemistry, 120(6), 10787-10795.

CAR Construction : 3D8 scFv-28ζ

Fig.8 Recognition of PSMA as target cells.

After 24 hours of coculturing T cells with target cells (PSMA+ LNCaP and PSMA− DU-145 cells at E:T ratios 1:1 and 3:1.

Hassani, M., Hajari Taheri, F., Sharifzadeh, Z., Arashkia, A., Hadjati, J., van Weerden, W. M., ... & Abolhassani, M. (2019). Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer. Journal of Cellular Biochemistry, 120(6), 10787-10795.

CAR Construction : 3D8 scFv-28ζ

Fig.9 Proliferation of engineered T cells upon encountering PSMA.

Data are normalized with the growth rate of negative control cell and reported as a percentage.

Hassani, M., Hajari Taheri, F., Sharifzadeh, Z., Arashkia, A., Hadjati, J., van Weerden, W. M., ... & Abolhassani, M. (2019). Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer. Journal of Cellular Biochemistry, 120(6), 10787-10795.