These data show that the dose-effect relationship between pfu/ml Ad5- 
CMV-lacZ concentrations and lacZ expression is similar for mouse nasal and 
human nasal epithelia. As in human tissues, effects of adenovirus on cell 
morphology are not observed. 
IYA.2.a.(ii) Xenograft model 
For these studies, a virus with a similar overall structure to that of the Ad5- 
CB-CFTR and Ad5-CMV-lacZ viruses was utilized. This recombinant adenovirus, 
called Ad5-RSV-lacZ, has a lacZ minigene expressed from a RSV promoter in 
place of El sequences as well as deletion of the E3 region. Into the lumen of the 
grafts were injected the following viruses: 1) Ad5-RSV-lacZ, 2) El deleted 
adenovirus (AdEldelta), 3) wild type Ad5, and 4) PBS ("mock"). The virus was 
expelled and the xenografts were subjected to the analyses summarized in Appendix 
E. Specific details of the experiments are provided below. 
IVA.2.a.(ii)(a) Efficiency of adenoviral-mediated gene 
transfer in human bronchial epithelium 
Xenografts were exposed to concentrated purified stocks of Ad5-RSV-lacZ 
adenovirus at 10^ pfu/ml, l(r pfu/ml, and 10^ pfu/ml for a total of 1 hour. The 
El deleted adenovirus (AdEldelta) at lO 1 ^ pfu/ml was used as a negative control. 
The grafts were harvested, fixed, stained in X-gal, and visualized enface through a 
dissecting microscope in order to assess the overall efficiency of infection (Figure 5). 
X-gal cytochemical analysis of adenovirally infected xenografts. Whole mount photomicrographs of 
X-gal stained bronchial xenografts infected with Ad5-RSV-lacZ at 10^ pfu/ml (A), 10^ (B and C), 
and 10 12 pfu/ml (D). The inset in (D) represents a 3 day old graft infected with AdEldelta. All 
xenografts were harvested at 3 days postinfection except for (C), which was harvested 21 days after 
infusion of virus. Long arrows point to "clonal" aggregates of lacZ positive cells while short arrows 
point to single isolated infected cells (bar = 400 jLlm). 
Figure 5. Ad5-RSV-lacZ exposed human airway epithelial xenografts. 
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Recombinant DNA Research, Volume 17 
