Extended Data Fig. 10: BCMA/SLAMF7 ChTCR constructs, antigen sensitivity and signaling comparison of BCMA/SLAMF7 Bi-ChTCR T cells.
a. Schematic of SLAMF7- and BCMA- mono- and bispecific receptor constructs. b. Absolute numbers ± SD of BCMA and SLAMF7 CAR and Bi-ChTCR T cells over 10 days of culture. (n = 3 independent experiments) c. and d. INF-γ production (pg/mL) of CAR and Bi-ChTCR in response to various concentrations of plate bound recombinant BCMA protein (c) or recombinant SLAMF7 protein (d) (n = 3 independent experiments). e. Representative flow plots showing binding of recombinant BCMA-APC and SLAMF7-PE to primary CD8 T cells transduced with the BCMA/SLAMF7 bispecific CAR or Bi-ChTCR. f. and g. Geometric mean of rBCMA-APC (f) and rSLAMF7-PE binding to bispecific CAR and bi-ChTCR T cells (g) (gated on receptor positive cells; n = 2 independent experiments). h. Representative western blot analysis for pZap70 pTyr319 and Zap70 for Bi-ChTCR or bispecific CAR stimulated with 0.5 M of rBCMA (1), 0.5 M of rSLAMF7 (2), 0.25 M of rBCMA + 0.25 M rSLAMF7 (3) or .05 M rBCMA + 0.5 M rSLAMF7 (4) coated-beads for indicated time. i. Mean band intensity for pZap70 pTyr319 normalized to total Zap70 (n = 2 independent experiments).
