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. 2022 Aug 18;2(8):827–841. doi: 10.1158/2767-9764.CRC-21-0154

FIGURE 1.

FIGURE 1

Development of a TCR-like CAR format (TCAR) that mediates antigen recognition and CD3 recruitment. A, Schematics of four TCAR formats designed by fusing CLDN6-specific antibody variable domains VH and VL in different orientations to either mouse or human TCR constant domains. The membrane-proximal native and the distal artificial disulfide bonds are indicated. B, Surface expression (top) and antigen-specific stimulation (bottom) of different TCARs after RNA transfer in CD8+ T cells. TCAR surface expression was assessed by flow cytometry. Cells were gated on control CD8+ T cells and dashed line indicates threshold for gating of CAR+/CD8+ T cells. Antigen-specific stimulation of TCAR T cells was evaluated by IFNγ ELISA (E:T = 10:1) after 20 hours of coculture with CLDN6+ DCs. T cells transfected with prototype β-chain RNA only (Ctrl.Tc) and DCs transfected with gp100 RNA (Ctrl. RNA) were used as controls. Graph shows mean + SD of technical duplicates. C, Surface expression of CD3ε with different TCAR formats, tested in Jurkat76 cells, which lack endogenous TCR and therefore surface expression of CD3. Murine CLDN6-TCR (TCR), CLDN6-CAR-28ζ, and TCAR prototype α-chain only (Ctrl.Tc) were used as controls.