Grafts exposed to the lacZ virus demonstrated autofluorescence below the 
basal laminal and low level fluorescence in basal cells representing endogenous 
CFTR expression (panels A and B). The grafts exposed to Ad5-CB-CFTR 
demonstrated a high level of CFTR expression in approximately 2-10% of the cells. 
Both ciliated and secretory cells expressed the recombinant CFTR, which in each 
case was localized on the apical basal lamina. Recombinant CFTR was also 
expressed in a number of intermediate cells. All types of surface epithelial cells 
expressed the recombinant CFTR (panels C and D). These experiments indicate 
that high levels of recombinant CFTR expression can be achieved in the surface 
epithelium. Furthermore, the recombinant protein is appropriately localized to the 
apical plasma membrane. 
IVA.2.a.(ii)(b) Efficiency of infection of the various 
bronchial epithelial cell types 
Quantification of the various cell types infected by 10^ pfu/ml adenovirus 
was performed by analysis of 1,000 transgene-expressing cells detected by X-gal 
staining and (3-galactosidase immunocytochemistry. Figure 8 shows the distribution 
of X-gal positive cell types as compared to the normal distribution of that cell type 
within the epithelium. 
cilated basal goblet intermediate 
cell type 
Figure 8. Distribution of cell types (solid bar) and transduced cell types (hatched 
bar) in xenografts. 
With a total of 11.1% lacZ positive cells in the epithelium, 5.5% were 
ciliated cells, 0.3% were basal cells, 3.8% were goblet cells, and 1.5% were 
intermediate cells. The diffusion of X-gal precipitate and the apparent non-nuclear 
localization of a majority of the expressing cells made the quantification of cell types 
within these large, highly expressing clusters difficult. We performed quantitation of 
lacZ positive cells by immunocytochemistry using a (S-galactosidase polyclonal 
[462] 
Recombinant DNA Research, Volume 17 
