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. 2023 Feb 7;29(12):2250–2265. doi: 10.1158/1078-0432.CCR-22-3298

Figure 5.

Figure 5. Evaluation of the tumor specificity of the three immunogenic nonC-TL. A, RNA expression analysis in tumors (T) and matched healthy tissues (N) of the canonical genes encoding immunogenic nonC-TL compared with TAA and CGA. TCGA and GTEX data were obtained from GEPIA. B, CD8 coreceptor activation-dependence of PBL transduced with antigen-specific TCRs. FACS plots show the expression of 4–1BB by CD8 (gated on CD3+mTCR+) after coculture with peptide pulsed B cells. B cells pulsed with an irrelevant (Irrel.) peptide were used as negative control. C, TCR-transduced T cells purified by FACS sorting (CD8+mTCR+) were cocultured with B cells pulsed with serial dilutions of the corresponding peptide. SD mean is plotted. D, Expression and translation of nonC-TL in healthy human cells and selected TCLs was indirectly evaluated by coculturing control and electroporated target cells with RNA encoding the specified HLA alleles with sort purified TCR-transduced T cells. T-cell activation was assessed by measuring 4–1BB expression on CD8+mTCR+. ¥TCL naturally expressing HLA-A*11:01. n.a, non-assessed.

Evaluation of the tumor specificity of the three immunogenic nonC-TL. A, RNA expression analysis in tumors (T) and matched healthy tissues (N) of the canonical genes encoding immunogenic nonC-TL compared with TAA and CGA. TCGA and GTEX data were obtained from GEPIA. B, CD8 coreceptor activation-dependence of PBL transduced with antigen-specific TCRs. FACS plots show the expression of 4–1BB by CD8 (gated on CD3+mTCR+) after coculture with peptide pulsed B cells. B cells pulsed with an irrelevant (Irrel.) peptide were used as negative control. C, TCR-transduced T cells purified by FACS sorting (CD8+mTCR+) were cocultured with B cells pulsed with serial dilutions of the corresponding peptide. SD mean is plotted. D, Expression and translation of nonC-TL in healthy human cells and selected TCLs was indirectly evaluated by coculturing control and electroporated target cells with RNA encoding the specified HLA alleles with sort purified TCR-transduced T cells. T-cell activation was assessed by measuring 4–1BB expression on CD8+mTCR+. ¥TCL naturally expressing HLA-A*11:01. n.a, non-assessed.