All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.
The vector of anti-CD20 chimeric antigen receptor(CAR) is constructed for the engineering of NK cells to target Human CD20. The NK cells are genetically modified through transduction with a Lentiviral vector expressing scFv of anti-CD20 antibody linked to CD28 transmembrane domain/ endodomain and CD137 (4-1BB), CD3-zeta signaling domains. And the vector product was designed for the treatment of chronic lymphocytic leukemia (CLL).
CAR Construction : rituximab scFv-41BB-CD28ζ Fig.1 Rituximab blocks antigen binding of Ab used to derive CAR scFv. Ramos cells (CD20þ) were incubated with the indicated rituximab concentrations (conc.) for 30 minutes, followed by incubation with PE-labeled anti-CD20 Ab (clone Leu16) or isotype control at either 4C or 37C for 30 minutes. Cells were washed and analyzed by flow cytometry to determine available CD20 binding sites as measured by PE fluorescence intensity. The panel summarizes the geometric mean fluorescence intensity (MFI) at either 4C or 37C as a function of rituximab concentration. Rufener, G. A., Press, O. W., Olsen, P., Lee, S. Y., Jensen, M. C., Gopal, A. K., Pender, B., Budde, L. E., Rossow, J. K., Green, D. J., Maloney, D. G., Riddell, S. R., & Till, B. G. (2016). Preserved Activity of CD20-Specific Chimeric Antigen Receptor-Expressing T Cells in the Presence of Rituximab. Cancer immunology research, 4(6), 509–519. |
CAR Construction : rituximab scFv-41BB-CD28ζ Fig.2 Effect of rituximab on CAR T-cell function in vitro. The indicated B-cell NHL cell lines were irradiated and incubated for 30 minutes at room temperature with varying rituximab concentrations (at two times the concentrations during incubation to yield the indicated final concentrations after addition of T cells). Rufener, G. A., Press, O. W., Olsen, P., Lee, S. Y., Jensen, M. C., Gopal, A. K., Pender, B., Budde, L. E., Rossow, J. K., Green, D. J., Maloney, D. G., Riddell, S. R., & Till, B. G. (2016). Preserved Activity of CD20-Specific Chimeric Antigen Receptor-Expressing T Cells in the Presence of Rituximab. Cancer immunology research, 4(6), 509–519. |
CAR Construction : rituximab scFv-41BB-CD28ζ Fig.3 Effect of rituximab on CAR T-cell– mediated cytotoxicity. The indicated 51Cr-labeled target cells were preincubated for 30 minutes with rituximab (at two times the concentrations during incubation Rufener, G. A., Press, O. W., Olsen, P., Lee, S. Y., Jensen, M. C., Gopal, A. K., Pender, B., Budde, L. E., Rossow, J. K., Green, D. J., Maloney, D. G., Riddell, S. R., & Till, B. G. (2016). Preserved Activity of CD20-Specific Chimeric Antigen Receptor-Expressing T Cells in the Presence of Rituximab. Cancer immunology research, 4(6), 509–519. |
CAR Construction : rituximab scFv-41BB-CD28ζ Fig.4 In vivo effect of rituximab on CD20 CAR T-cell function. NSG mice were injected i.v. with 5 105 rituximab-refractory Raji-ffLuc lymphoma cells, followed by one of the following treatments: no treatment, rituximab only (25 mg or 200 mg) i.p. 5 days later, 107 1.5.3-NQ-28-BBz CAR T cells only 6 days after tumor, |
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