FIG. 8.

In vivo biodistribution. hCD46Ge mice were injected in the tail vein with 10¹¹ viral particles of Ad5-CMV-GFP, Ad5/11-CMV-GFP, and Ad11-CMV-GFP. Organs were obtained from individual mice at 30 min and 72 h postinjection. (a) Tissue biodistribution of vector genomes. Genomic DNA was isolated from organs harvested at 30 min and 72 h postinjection, and the presence of Ad vector genomes in liver, lung, kidney, spleen, and bone marrow was analyzed by Southern blotting with a GFP-specific probe. Membranes were stripped of GFP probe and rehybridized with a housekeeping mouse β-glucoronidase (GUS)-specific probe. (b) In vivo gene transfer. Vector mediated expression of GFP in liver harvested 72 h postinjection was detected with a monoclonal anti-GFP antibody. Binding of primary antibody was visualized with fluorescein isothiocyanate-conjugated secondary antibody and a fluorescence microscope (green) or with horseradish peroxidase-conjugated secondary antibody and diaminobenzidine tetrahydrochloride (brown). Arrows indicate GFP-positive nonparenchymal liver cells. Magnification, ×20 (left three images) and ×40 (right image).