Figure 3
Example of co-stimulatory molecule expression by melanoma cells after transduction with TFG tricistronic vector: Immunostaining with hCD80 –(x-axis) and hCD70 – (y-axis) 3A: non transduced melanoma cells were cultured. Ccells (3 × 105) was stained for surface expression of CD70 and CD80 using specific antibodies as described in the Methods. 3B-3E: Murine B16.F10 melanoma cells were transduced with the four TFG retroviral vectors. Forty eight hours after transduction, pools of cells (3 × 105) were stained for surface expression of CD70 and CD80 using specific antibodies as described in the Methods. 2B: TFGEMCVZEO transduced cells, 2C: TFGc-MYCZEO transduced cells, 2D: TFGFGF-2ZEO transduced cells, 2E: TFGHTLV-1ZEO transduced cells.3F: Human melanoma cells were transduced with the TFGFGF-2ZEO. A selected pool of cells (3 × 105) was stained for surface expression of CD70 and CD80 using specific antibodies as described in the Methods. The samples were subjected to two-color analysis by flow cytometry. Data are shown as immunofluorescence profiles with arbitrary fluorescence units (log) of FITC on the x-axis and immunofluorescence profiles with arbitrary fluorescence units (log) of PE on the y-axis. To evaluate the percentage of co-stimulatory molecules expressed by transfected cells, the marker was set to allow < 5% positive cells in the non-transduced cells.