Figure 4.

Isolation of a Mutated NBAS-Specific TCR

(A and B) TIL4112F5 T cells were co-cultured with TMG-9-transfected autologous DCs for 4 hr, and single-cell RNA-seq was performed. The expression of IFN-γ and IL-2 mRNA of individual single cells is shown in dot plots (A) and the scatter plot (B). (C) 4112TCR was transduced into donor T cells, and then transduced T cells were co-cultured with TMG-transfected autologous DCs overnight. Error bars represent SD. (D) 67 predicted short peptides were combined into 10 short-peptide pools (SPPs) and pulsed on autologous EBV-transformed B cells for 2 hr. Peptide-pulsed EBV-transformed B cells were then co-cultured with 4112TCR-transduced T cells overnight. (E) 7 predicted short peptides from SPP-9 were individually pulsed on autologous EBV-transformed B cells for 2 hr. Peptide-pulsed EBV-transformed B cells were then co-cultured with 4112TCR-transduced T cells overnight. (F) Purified mutated NBAS peptide (WSYDSTLLAY, C > S) or its WT counterpart was pulsed on autologous EBV-transformed B cells for 2 hr, and then peptide-pulsed B cells were co-cultured with 4112TCR-transduced T cells overnight. The secretion of IFN-γ from T cells was determined by ELISA.