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. 2022 Feb 20;24:729–741. doi: 10.1016/j.omto.2022.02.012

Figure 2.

Figure 2

Functional characterization of the novel bispecific IL13 (4MS)-EphA2 scFv-TanCAR in Jurkat T cells

(A) Cartoon of the novel bispecific IL13 (4MS)-EphA2 scFv-TanCAR. To construct this novel TanCAR, IL13 (4MS) was inserted into a third-generation CAR backbone, then EphA2 scFv was linked with IL13 (4MS) by a (GGGS)3 linker. (B) Illustration of a cell-based luciferase reporter system for analyzing the biological activity of the novel TanCAR. (C) The biological activity assay of IL13 (4MS)-EphA2 scFv-TanCAR with U87 cells using a cell-based luciferase reporter system. (D) The biological activity assay of IL13 (4MS) CAR, EphA2 scFv CAR or IL13 (4MS)-EphA2 scFv-TanCAR with U87 cells using a cell-based luciferase reporter system. (E) The biological activity assay of IL13 (4MS) CAR, EphA2 scFv CAR, or IL13 (4MS)-EphA2 scFv-TanCAR with EphA2-engineered K562 target cells, IL13Rα2-engineered K562 target cells, or EphA2-IL13Rα2-engineered K562 target cells individually using a cell-based luciferase reporter system. Shown are representative plots of at three independent experiments performed in triplicate. Statistically significant differences are indicated: ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. TanCAR stands for IL13 (4MS)-EphA2 scFv-TanCAR.