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The vector of anti-EGFRvIII chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human EGFRvIII. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-EGFRvIII antibody linked to OX40 and CD3ζ signaling domains. And the vector product was designed for the treatment of Human glioblastoma (GBM).
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CAR Construction :
Fig.1 NIH-3T3 cells were transduced with a retroviral vector expressing EGFRvIII and a neomycin resistance gene. Following selection in neomycin analogue G418, cells were subject to FACS analysis with anti-EGFRvIII mAb as shown. Morgan, R. A., Johnson, L. A., Davis, J. L., Zheng, Z., Woolard, K. D., Reap, E. A., ... & Rosenberg, S. A. (2012). Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma. Human gene therapy, 23(10), 1043-1053. |
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CAR Construction : 3C10 scFv-28ζ
Fig.2 Development of CARs targeting EGFRvIII. scFv genes were synthesized and fused to T-cell signaling domains from CD28 and CD3f followed by insertion into c-retroviral vector MSGV1 (diagram). Morgan, R. A., Johnson, L. A., Davis, J. L., Zheng, Z., Woolard, K. D., Reap, E. A., ... & Rosenberg, S. A. (2012). Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma. Human gene therapy, 23(10), 1043-1053. |
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CAR Construction : 3C10 scFv-28ζ
Fig.3 Anti-EGFRvIII CAR vector-engineered T cells specifically recognize EGFRvIII-expressing cells. c-Retroviral vectors expressing the anti-EGFRvIII CARs (from murine mAbs 3C10 and L8A4 and human mAb 139) were used to transduced human T cells. Morgan, R. A., Johnson, L. A., Davis, J. L., Zheng, Z., Woolard, K. D., Reap, E. A., ... & Rosenberg, S. A. (2012). Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma. Human gene therapy, 23(10), 1043-1053. |
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