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. 2022 Jul 25;21(10):1608–1621. doi: 10.1158/1535-7163.MCT-22-0059

Figure 4.

Figure 4. Tuning anti-FGFR4 CARs for low-density antigens. Expression of RH30_19 antigen surface expression of FGFR4 and CD19 as assessed by flow cytometry (A) and quantified with QuantiBrite beads (B). Quantification of surface FGFR4 expression of cultured RH30_19 and RD cell lines or RH30_19 cells derived from excised tumors. Both flow analysis (left) and quantification (right) are shown. C, New CAR structural and signaling formats evaluated: Double CD3ζ CAR, and CD28 H/TM CAR. D, Surface expression of RJ154HL CAR (anti-FGFR4 CAR) with indicated new signaling domains assessed by flow cytometry. E, Cytotoxicity of anti-FGFR4 CAR T cells against the RD-ffLuc RMS cell line after 5 hours of co-culture at indicated E:T ratios. F, Cytokine production by anti-FGFR4 CAR after 20 hours of co-culture with RH30_19 target cells. Average of three wells and SEM is indicated for each new CAR format and control UTD.

Tuning anti-FGFR4 CARs for low-density antigens. Expression of RH30_19 antigen surface expression of FGFR4 and CD19 as assessed by flow cytometry (A) and quantified with QuantiBrite beads (B). Quantification of surface FGFR4 expression of cultured RH30_19 and RD cell lines or RH30_19 cells derived from excised tumors. Both flow analysis (left) and quantification (right) are shown. C, New CAR structural and signaling formats evaluated: double CD3ζ CAR, and CD28 H/TM CAR. D, Surface expression of RJ154HL CAR (anti-FGFR4 CAR) with indicated new signaling domains assessed by flow cytometry. E, Cytotoxicity of anti-FGFR4 CAR T cells against the RD-ffLuc RMS cell line after 5 hours of coculture at indicated E:T ratios. F, Cytokine production by anti-FGFR4 CAR after 20 hours of coculture with RH30_19 target cells. Average of three wells and SEM is indicated for each new CAR format and control UTD.