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The vector of anti-Lewis Y chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Human Lewis Y. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-Lewis Y antibody linked to 4-1BB and CD3ζ signaling domains. And the vector product was designed for the treatment of gastric cancer.
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CAR Construction :
Fig.2 Flow cytometric analysis of binding activity of mAbPand mAbMBR55-2 to the FcRI receptor. U937 cells treated with IFN to stimulate Fc receptorexpression were incubated with mAbPor mAbMBR55-2 or CD64-specific mAb m22-FITC. Brodzik, R., Glogowska, M., Bandurska, K., Okulicz, M., Deka, D., Ko, K., ... & Koprowski, H. (2006). Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells. Proceedings of the National Academy of Sciences, 103(23), 8804-8809. |
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CAR Construction :
Fig.3 ELISA plates were coated overnight with the peptides, and reactivity of serial dilutions of the mAbs was detected. Plates were then washed and incubated with post-immune serum from a patient immunized with conjugated P10s. Nounamo, B., Jousheghany, F., Siegel, E. R., Post, S. R., Kelly, T., Ferrone, S., ... & Monzavi-Karbassi, B. (2023). VT68. 2: An Antibody to Chondroitin Sulfate Proteoglycan 4 (CSPG4) Displays Reactivity against a Tumor-Associated Carbohydrate Antigen. International Journal of Molecular Sciences, 24(3), 2506. |
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CAR Construction :
Fig.1 MDA-MB-231 cells were preincubated with Caspase-3 Inhibitor Z-DEVD-FMK or DMSO (control) for 2 hours and then pre- and post-immune plasma were added and incubation continued overnight. Cells then were harvested and stained with annexin V-FITC (FL1) and propidium iodide (FL3) using a live/dead assay kit. Hutchins, L. F., Makhoul, I., Emanuel, P. D., Pennisi, A., Siegel, E. R., Jousheghany, F., ... & Kieber-Emmons, T. (2017). Targeting tumor-associated carbohydrate antigens: a phase I study of a carbohydrate mimetic-peptide vaccine in stage IV breast cancer subjects. Oncotarget, 8(58), 99161. |
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