Anti-CEA (hMN14) h(CD28-CD3ζ) CAR, pCDCAR1 (CAR-ZP041)

CAR Construction :

Fig.1 Identifying MN-14 epitope-containing domain of CEA. FACS profiles of transfectant cells were obtained after staining with humanized anti-CEA Ab hMN-14.

All other constructs were expressed in LR-73 cells that have normal glycosylation.

Ma, Q., DeMarte, L., Wang, Y., Stanners, C. P., & Junghans, R. P. (2004). Carcinoembryonic antigen-immunoglobulin Fc fusion protein (CEA-Fc) for identification and activation of anti-CEA immunoglobulin-T-cell receptor-modified T cells, representative of a new class of Ig fusion proteins. Cancer gene therapy, 11(4), 297-306.

CAR Construction :

Fig.2 Identification of MN-14 antibody and Fig.3 Anti-CEA IgTCR by CEA-Fc.

Anti-CEA IgTCR chimeric receptor was detected by flow cytometric analysis using CEA-Fc.

Ma, Q., DeMarte, L., Wang, Y., Stanners, C. P., & Junghans, R. P. (2004). Carcinoembryonic antigen-immunoglobulin Fc fusion protein (CEA-Fc) for identification and activation of anti-CEA immunoglobulin-T-cell receptor-modified T cells, representative of a new class of Ig fusion proteins. Cancer gene therapy, 11(4), 297-306.

CAR Construction :

Fig.3 Activation of anti-CEA IgTCR-transduced Jurkat T cells by CEA-Fc.

Jurkat and anti-CEA IgTCR-transduced Jurkat cells (Jurkat-IgTCR) were incubated for 24 hour with BSA, OKT3, CEA, and CEA-Fc in soluble form.

Ma, Q., DeMarte, L., Wang, Y., Stanners, C. P., & Junghans, R. P. (2004). Carcinoembryonic antigen-immunoglobulin Fc fusion protein (CEA-Fc) for identification and activation of anti-CEA immunoglobulin-T-cell receptor-modified T cells, representative of a new class of Ig fusion proteins. Cancer gene therapy, 11(4), 297-306.

CAR Construction : CEA scFv-TM28-CD28-CD3ζ

Fig.4 CD8+ T cells were transduced with the retroviral constructs and GFP+ CD8+ T cells (upper panel) or GFP+ Myc+ T cells gated on GFP+ CD8+ populations were analyzed by flow cytometry

In vitro generation of carcinoembryonic antigen (CEA) chimeric antigen receptor T (CAR-T) cells.

Fan, J., Das, J. K., Xiong, X., Chen, H., & Song, J. (2021). Development of CAR-T cell persistence in adoptive immunotherapy of solid tumors. Frontiers in oncology, 10, 574860.

CAR Construction : CEA scFv-TM28-CD28-CD3ζ

Fig.5 The packaging Plat-E cells (upper panel) and MFG-transduced CD8+ T cells (lower panel) were visualized by fluorescence microscope.

scale bars: 200 mm

Fan, J., Das, J. K., Xiong, X., Chen, H., & Song, J. (2021). Development of CAR-T cell persistence in adoptive immunotherapy of solid tumors. Frontiers in oncology, 10, 574860.

CAR Construction : CEA scFv-TM28-CD28-CD3ζ

Fig.6 GFP+ CD8+ T cells were sorted, and the cell lysates were determined for the expression of Bcl-xL and b-actin by western blotting.

All data are representative of three independent experiments.

Fan, J., Das, J. K., Xiong, X., Chen, H., & Song, J. (2021). Development of CAR-T cell persistence in adoptive immunotherapy of solid tumors. Frontiers in oncology, 10, 574860.

CAR Construction : CEA scFv-TM28-CD28-CD3ζ

Fig.7 Carcinoembryonic antigen (CEA) chimeric antigen receptor T (CAR-T) cells overexpressing Bcl-xL had modest in vitro cytotoxicity.

Representatives of the in vitro co-culture of T cells with MC-32 tumor cells at 12 h (scale bars: 50 mm).

Fan, J., Das, J. K., Xiong, X., Chen, H., & Song, J. (2021). Development of CAR-T cell persistence in adoptive immunotherapy of solid tumors. Frontiers in oncology, 10, 574860.

CAR Construction : CEA scFv-TM28-CD28-CD3ζ

Fig.8 Representatives of the CD8+ Thy1.2+ T cells in the LNs and spleen on day 14.

All data are representative of three independent experiments.

Fan, J., Das, J. K., Xiong, X., Chen, H., & Song, J. (2021). Development of CAR-T cell persistence in adoptive immunotherapy of solid tumors. Frontiers in oncology, 10, 574860.