Figure 2

Transduction with Ad5.UbP.GFP and Ad5.CMV5.GFP of lymph node cells and thymocytes from line 18 mice. Lymph node cells and thymocytes from line 18 mice were isolated and transduced with Ad5.UbP.GFP or Ad5.CMV5.GFP adenoviral reporter vectors at a MOI of 10 for 30' and incubated for 24 h at 37°C/5%CO₂. Half of the cells transduced with Ad5.CMV5.GFP were stimulated for 4 h with PMA/ionomycin before analysis. Lymph node cells were then stained with anti-CD4, anti-CD25 and anti-CD62L; thymocytes were stained with anti-CD4 and anti-CD8. The quadrants were set based on the transduction of hCARΔcyt negative littermate controls (data not shown). A. Two color dot-plots of lymph nodes cells transduced with Ad5.Ubp.GFP or Ad5.CMV5.GFP with or without activation with PMA/ionomycin. The top panel represent the expression of hCARΔcyt and CD4 detected in the viable lymphoid gate while the two bottom panel were gated on CD4 positive cells only (rectangular gate). The percentage of cells in each quadrant is indicated. B. Two-color dot plot of CD4 and CD8 expression of line 18 thymocytes transduced with Ad5.UbP.GFP. The histograms represent the GFP expression for each thymocyte subset defined by their CD4 and CD8 expression. The percentage of GFP positive cells is indicated.