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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 and CD3ζ signaling domains. And the vector product was designed for the treatment of Glioma.
CAR Construction : MAb 139 scFv-28ζ Fig.1 EGFRvIII mCAR retroviral gene delivery to murine T cells confers antigen-specific activity. VM/Dk-derived glioma SMA560vIII shows EGFRvIII expression compared with SMA560 by surface antibody staining (L8A4) and flow cytometry. Sampson, J. H., Choi, B. D., Sanchez-Perez, L., Suryadevara, C. M., Snyder, D. J., Flores, C. T., ... & Johnson, L. A. (2014). EGFRvIII mCAR-Modified T-Cell Therapy Cures Mice with Established Intracerebral Glioma and Generates Host Immunity against Tumor-Antigen LossEGFRvIII mCARs for Malignant Glioma. Clinical Cancer Research, 20(4), 972-984. |
CAR Construction : MAb 139 scFv-28ζ Fig.2 EGFRvIII mCAR T-cell therapy against intracerebral tumors requires lymphodepletive host-conditioning for efficacy, and can be abrogated by soluble peptide administration. To evaluate the impact of lymphopenia on the antitumor efficacy of EGFRvIII mCAR T cells, VM/Dk mice (8 per group) with 3-day established intracerebral SMA560vIII tumors. Sampson, J. H., Choi, B. D., Sanchez-Perez, L., Suryadevara, C. M., Snyder, D. J., Flores, C. T., ... & Johnson, L. A. (2014). EGFRvIII mCAR-Modified T-Cell Therapy Cures Mice with Established Intracerebral Glioma and Generates Host Immunity against Tumor-Antigen LossEGFRvIII mCARs for Malignant Glioma. Clinical Cancer Research, 20(4), 972-984. |
CAR Construction : MAb 139 scFv-28ζ Fig.3 Treatment of mice bearing EGFRvIIIPOS tumors with mCARs provides long-term protection against EGFRvIIINEG tumor SMA560 (EGFRvIIINEG) gliomas were maintained in vitro or passaged in a mouse subcutaneously for 14 days, then harvested and both were stained for EGFRvIII expression using L8A4 mAb and analyzed by flow cytometry. Sampson, J. H., Choi, B. D., Sanchez-Perez, L., Suryadevara, C. M., Snyder, D. J., Flores, C. T., ... & Johnson, L. A. (2014). EGFRvIII mCAR-Modified T-Cell Therapy Cures Mice with Established Intracerebral Glioma and Generates Host Immunity against Tumor-Antigen LossEGFRvIII mCARs for Malignant Glioma. Clinical Cancer Research, 20(4), 972-984. |
CAR Construction : MAb 139 scFv-28ζ Fig.4 Development of CARs targeting EGFRvIII. NIH-3T3 cells and BHK cells were transduced with a retroviral vector expressing EGFRvIII and a neomycin resistance gene. Based on the amino acid sequence for anti-EGFRvIII mAbs, 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. |
CAR Construction : MAb 139 scFv-28ζ Fig.5 Equivalent function of second- and third-generation anti-EGFRvIII CAR vectors. Using anti-EGFRvIII human mAb 139, a third-generation CAR vector was assembled using CD28, 4-1BB, and CD3f T-cell signaling domains. 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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