Figure 1

Flow cytometry to detect Ad attachment to nucleated human cells. (a) One million cells of the isogenic human melanoma cell lines Mo and the stably-transfected Mo-αVβ3 and Mo-αIIbβ3 cell lines (respectively expressing αVβ3 integrin and the platelet αIIbβ3 integrin) were incubated with Ad (MOI = 10, 4°C, 1-hr), followed by rinse and staining with a FITC-labeled anti-Ad hexon antibody. The negative control comprised omitting Ad. Histograms show the distribution and fluorescence intensity of Ad bound to the cell surface (b) Ad infection in the above cell lines was studied using a replication deficient Ad vector expressing GFP (AdGFP). Cells were incubated with AdGFP at an MOI of 10 for 4 hours at 37°C, medium replaced and cells further cultured for 18-hrs. Intracellular GFP expression was measured using flow cytometry. *, p < 0.05 for enhanced Ad infection of Mo-αVβ3 vs. Mo cells and Mo vs. Mo-αIIbβ3 cell. Representative images of at least 2 different experiments (n = 3 for each).