Table 2. PBMC stimulation with hIFN-γ+GGTI-298 pretreated LB1319-MEL cells induces CD8+ and MART-1/HLA-A0201+ specific cells proliferation.
Percentage (%) of cells CD8+ and MART1/HLA-A2+ | Fold Induction (FI) of cells CD8+ and MART1/HLA-A2+ | |||||
HD1 | HD2 | HD3 | HD1 | HD2 | HD3 | |
NT | 1,5 | 0,29 | 0,07 | 1 | 1 | 1 |
IFN | 2,7 | 0,39 | 0,11 | 1,8 | 1,34 | 1,57 |
GGTI | 1,03 | 0,65 | 0,03 | 0,68 | 2,24 | 0,42 |
IFN+GGTI | 4,8 | 1,13 | 0,35 | 3,2 | 3,9 | 5 |
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.