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. 2021 Oct 9;9(10):1422. doi: 10.3390/biomedicines9101422

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Manufactured with G-Rex system, bispecific CS1-BCMA CAR-T cells express high functional activity with CS1 and BCMA-positive target cells. (A) FACS staining detects CS1 scFv and BCMA scFv-positive CAR-T cells. FACS was performed on T cells (top panel) and CS1-BCMA CAR-T cells (bottom panel) with biotinylated recombinant BCMA (left) and CS1 proteins (middle) and mouse F(ab’)2 antibody (right). (B,C) The percentage of CD4+/CD8+ cells is shown in the left panel, and the percentage of cells with a T_N, T_CM, T_EM, and T_EF phenotype is shown in the right panel. (C) Manufactured CS1-BCMA CAR-T cells secreted IFN-gamma with Hela-BCMA target cells. (D) CS1-BCMA CAR-T cells secreted IFN-gamma with CHO-CS1-positive target cells. (E) CS1-BCMA-CAR-T cells had significantly higher levels of IFN-gamma with BCMA/CS1-positive target cells than with T and Mock-CAR-T cells: Raji cells are BCMA-positive cells and MM1S cells are CS1-BCMA-positive cells. (C–E) p < 0.05, Student’s t-test, CS1-BCMA CAR-T cells versus Mock CAR-T cells.