Fig. 4.

Specificity of tumor-reactive T cells enriched in DT MLTC. Independent autologous MLTC were performed with PBMC collected from patient DT during a 4-year period (Fig. 1). CD8⁺ MLTC responders were isolated and cryopreserved at various time points and later tested in 20-h IFN-γ ELISPOT assays for recognition of autologous melanoma cells (30,000 per well) (M) and COS-7 cells (20,000 per well) transiently transfected with expression plasmids encoding the antigen/HLA combinations tyrosinase/HLA-A^*2601 (A), tyrosinase/HLA-B^*3801 (B), gp100/HLA-B^*07021 (C), SIRT2_v3^mut/HLA-A^*03011 (D), GPNMB_v2^mut/HLA-A^*03011 (E), SNRP116^mut/HLA-A^*03011 (F), RBAF600^mut/HLA-B^*07021 (G), SNRPD1^mut/HLA-B^*3801 (H). Data are means of duplicates and represent the number of spot forming cells per 15,000 (day 25) or 10,000 (later than day 25) CD8⁺ MLTC responders.