These results indicate that single exposure of a CF epithelium to Ad.CB-CFTR results in 
detectable differences in the bioelectric properties consistent with partial correction of CFTR 
function. 
II. B. 7. Safety Studies in Trangenic Mice 
Important safety concerns of this protocol relate to the consequences of ectopic and/or 
unregulated CFTR expression in the human lung. For example, will there be deleterious effects 
of CFTR expression in cells of the lung that do not normally express this gene? In collaboration 
with Dr. Jeff Whitsett, at the University of Cincinnati Medical Center, we established 
transgenic mouse lines that express human CFTR under the control of transcriptional elements 
derived from the human surfactant protein C(SP-C) gene [Whitsett et al. t 1992]. 
The hCFTR mRNA was expressed in lungs and testes: in the lung, we found hCFTR mRNA in 
bronchiolar and alveolar epithelial cells and CFTR protein in respiratory and epithelial cells. 
While the level of expression of hCFTR mRNA varied, hCFTR mRNA and protein were detected in 
pulmonary epithelial cells of several lines at high levels. Lung weight, morphology, somatic 
growth and reproductive capacity were not altered by expression hCFTR in lung and testes of the 
transgenics. Our findings suggest ectopic and unregulated expression of human CFTR in lung 
epithelial cells may not be deleterious. A copy of the paper describing this finding is provided 
in Appendix F. 
II. B. 8. Safety and Feasibility Studies in Nonhuman Primates 
Experiments were performed in three Rhesus monkeys (Macaca mulatta) to further assess the 
feasibility and safety of gene therapy using El deleted adenoviruses. Each animal was 
anesthetized, intubated, and ~3 ml of purified virus was administered into the airway through 
the endotracheal tube while the animal was lying in the left lateral decubitus position. Two 
animals (9037 and 9650) received Ad.CMV-lacZ and one animal (8829) received Ad.CB-CFTR. 
The animals were monitored clinically and serial blood hematology/chemistry were evaluated. 
Animals 9037 and 8829 were necropsied 3 days after administration of virus; animal 9650 
will be necropsied 3 weeks post-infusion. A summary of the experiment is presented in the 
table below. 
Virus 
Animal 
Sex 
Wt (Rg) 
Iyp£ 
TotaUplu) 
9037 
Male 
3.6 
Ad.CMV-lacZ 
3x1 0 1 1 
8829 
Male 
4.2 
Ad.CB-CFTR 
3x1 0 1 1 
9650 
Male 
3.2 
Ad.CMV-lacZ 
3x1 0 1 1 
The animals tolerated the procedure well without obvious clinical sequalae. Necropsy 
evaluations of animals 9037 and 8829 three days after administration of virus failed to reveal 
pathology. Blood chemistry and hematology values remained within normal limits (see Tables 
in Appendix A). 
We have begun characterizing tissues from 9037 for evidence of adenovirus mediated gene 
transfer. Tissues from various organs were frozen, cryosectioned, mounted, and stained in X- 
gal. No staining was detected above background in brain, liver, spleen, kidney, heart, and 
testes. Significant staining was seen throughout the lung. Representative sections of lung are 
presented in Figure 16. There was patchy staining in proximal airway covering 1-2% of the 
[818] 
Recombinant DNA Research, Volume 16 
