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Recombinant Moloney murine leukemia virus (MMLV) retroviral vectors serve as efficient viral vector tools for introducing genes permanently into a wide variety of dividing cells, and provide an alternative to modify primary T cells. Creative biolabs has developed retroviral CAR vector pMMLV TAG72 (huCC49) h(28ζ), which is constructed for the engineering of T cells to target human TAG72. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-TAG72 antibody linked to a CD28 transmembrane domain and CD3-zeta signaling domains. And the vector product was designed for the treatment of Ovarian cancer.
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CAR Construction : huCC49 scFv-41BB-CD3ζ
Fig.1 TAG72-specific CAR T cells containing a 4-1BB intracellular co-stimulatory domain. Mock (untransduced) and TAG72-BBζ CAR T cells were evaluated by flow cytometry for CD19t expression to detect lentiviral transduction of CARs (left) or Protein L to detect the scFv (right). Murad, J. P., Kozlowska, A. K., Lee, H. J., Ramamurthy, M., Chang, W. C., Yazaki, P., Colcher, D., Shively, J., Cristea, M., Forman, S. J., & Priceman, S. J. (2018). Effective Targeting of TAG72+ Peritoneal Ovarian Tumors via Regional Delivery of CAR-Engineered T Cells. Frontiers in immunology, 9, 2268. |
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CAR Construction : huCC49 scFv-41BB-CD3ζ
Fig.2 TAG72-BBζ CAR T cells show antigen-dependent cytokine production and tumor killing in vitro. Quantification of tumor killing and (H) IFNγ production by TAG72-BBζ CAR T cells relative to Mock following a 72 h co-culture with freshly thawed OAS cells at an E:T ratio of 1:1. Murad, J. P., Kozlowska, A. K., Lee, H. J., Ramamurthy, M., Chang, W. C., Yazaki, P., Colcher, D., Shively, J., Cristea, M., Forman, S. J., & Priceman, S. J. (2018). Effective Targeting of TAG72+ Peritoneal Ovarian Tumors via Regional Delivery of CAR-Engineered T Cells. Frontiers in immunology, 9, 2268. |
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CAR Construction : huCC49 scFv-41BB-CD3ζ
Fig.3 Regional intraperitoneal delivery of TAG72-BBζ CAR T cells significantly reduces tumor burden and extends overall survival of OVCAR3 tumor-bearing mice. Kaplan-Meier survival for Mock and TAG72-BBζ CAR T cell treated mice. N ≥ 4 mice per group. Combined data from two independent experiments. Murad, J. P., Kozlowska, A. K., Lee, H. J., Ramamurthy, M., Chang, W. C., Yazaki, P., Colcher, D., Shively, J., Cristea, M., Forman, S. J., & Priceman, S. J. (2018). Effective Targeting of TAG72+ Peritoneal Ovarian Tumors via Regional Delivery of CAR-Engineered T Cells. Frontiers in immunology, 9, 2268. |
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