PBMC stimulation with hIFN-γ+GGTI-298 pretreated LB1319-MEL cells induces CD8+ and MART-1/HLA-A0201+ specific cells proliferation.

PBMC from 3 HLA-A0201 HD were stimulated twice in vitro with LB1319-MEL cells untreated (NT); or pretreated with 50 UI/mL hIFN-γ (IFN); 10 µM GGTI-298 (GGTI); or with a combination of 50 UI/mL hIFN-γ and 10 µM GGTI-298 (IFN+GGTI) as indicated. At the end of the culture period, PBMC were stained with FITC-conjugated anti-CD8 monoclonal antibody and with PE-conjugated MART-1/HLA-A2 tetramers. Percentages of double positive CD8+ and MART-1/HLA-A2 tetramers+ PBMC are shown. Fold induction between double positive CD8+ and MART-1/HLA-A2+ cells obtained after co-culture with untreated or pre-treated LB1319-MEL cells is shown. We defined the percentage of double positive cells obtained with the untreated melanoma cells (NT) as equal to 1 and compared this to the values obtained in the other culture conditions.