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. 2022 Feb 25;24:887–896. doi: 10.1016/j.omto.2022.02.024

Figure 1.

Figure 1

BiTE-T cells have decreased CD3/TCRαβ expression and alloreactivity

(A) CD3/TCRαβ expression on human T cells treated with human CD19 BiTE (blinatumomab) in the presence of 3T3.mCD19 (left) or 3T3.hCD19 cells (right). (B) CD3/TCRαβ expression on CD19 or Her2 BiTE-T cells. (C) T cell activation and cytokine production after TCRαβ stimulation. (D–G) GvHD risk evaluation of BiTE-T cells. Male NSG mice received cyclophosphamide (CTX) or whole-body irradiation (TBI) first and then 10 million T cells 1 day after. Mice were monitored for GvHD. (D) Study design. (E) Weight change over time. (F) NSG mice survival. (G) T cell persistence in peripheral blood. (H) CD3/TCRαβ expression on CD19 BiTE-T cells in peripheral blood of NSG mice without targets. (I) CD3/TCRαβ expression over time on Her2 BiTE-T cells in peripheral blood of NSG mice without targets. (J) CD3/TCRαβ expression on CD19 BiTE-T cells in serial-killing assay on CD19-expressing NALM6 cells. (K) CD3/TCRαβ expression on Her2 BiTE-T cells in serial killing assay on A375.Her2 cells. (L and M) CD3/TCRαβ expression on Her2-targeting T cells in vivo. NSG mice were subcutaneously injected with A375.Her2 cells and treated with Her2 BiTE-T or CAR-T cells 8–10 days later. (L) CD3/TCRαβ expression on donor T cells in peripheral blood 3 weeks after T cell injection. (M) CD3/TCRαβ expression of tumor infiltrating T cells 4 weeks after T cell injection in the A375.Her2 solid-tumor model. (N) CD3/TCRαβ expression on co-cultured T cells. BiTE-T or CAR-T cells from one healthy donor (as donor T cells) were co-cultured with activated T cells from another healthy donor (as recipient T cells) for 72 h and subjected to flow cytometry. DTC, donor T cells; RTC, recipient T cells. (O) CD3/TCRαβ expression over time on host and donor T cells in allogeneic MLR assay. CD19 BiTE-T or CAR-T cells were co-cultured with HLA-mismatched PBMCs at a 1:10 E:T ratio in 96-well plates, and CD3/TCRαβ expression on host and donor T cells were followed over time. Data are presented as means ± SD. ∗p<0.05; ∗∗p<0.01; ∗∗∗p<0.001; ∗∗∗∗p<0.0001; ns, not significant.