108 grafts from 18 non-CF bronchial samples and 104 grafts from 13 CF bronchial 
samples. 
The xenografts have been subjected to extensive morphometric analysis in 
order to evaluate the suitability of the model. Blocks of tissue from a non-CF and 
CF xenograft were carefully examined by transmission electron microscopy. Overall 
structure of the epithelium, distribution of cell types (i.e., ciliated, goblet, basal, and 
undifferentiated), and ultrastructural features of the various types of cells was 
determined. These findings were compared to similar analyses of native main stem 
bronchus (non-CF and CF) located adjacent to the area from which the cells 
destined for xenografts were removed. Electron micrographs of non-CF and CF 
xenografts and corresponding areas of adjacent airway showed no obvious 
differences in the overall structure or ultrastructural features of the various samples 
except that the native structure occasionally appeared taller. The distribution of cell 
types did not significantly differ between the non-CF bronchus and non-CF 
xenograft or the non-CF and CF xenograft. The CF bronchus had a significantly 
increased distribution of goblet cells when compared to the CF xenograft suggesting 
environmental stimuli may be responsible, in part, for goblet cell hyperplasia in CF 
(32% in bronchus vs. 24% in graft). 
A detailed description of the electrophysiologic properties of the non-CF and 
CF xenografts are presented in section IV.A.2.d. Manuscripts describing the use of 
the xenograft model are contained in Appendices C, D, and E. 
III. ADENOVIRUS CONSTRUCT 
III.A. Vector Construction 
IIIA.1. Construction of pAdBglll 
The purpose of this step was to construct a plasmid containing the 5’ portion 
of Ad 5 with deletion of El sequences and a unique cloning site. A series of figures 
(Figure 1) in Appendix F summarize this plasmid construction. As shown in Panel 
A, the plasmid pEHX-L3 contains sequences from Ad5 spanning map units 1 to 
16.1. pEHX-L3 was digested with EcoRI and Bglll and a 5.2 kb fragment isolated, 
which contains the adenoviral sequences from map units 9.2-16.1 and the plasmid 
backbone [derived from pAT 153 and described in Falck-Pedersen et al., 1989 (69)]. 
The adenoviral sequences from map unit 0-1 which contains the 5’ inverted terminal 
repeat, origin of replication and encapsidation signal were amplified from the 
original pEHX-L3 using PCR to insert a Nhe I site immediately downstream of the 
EcoR I site, and a Bgl II site at the 3’ end. This PCR fragment and the EcoR I/Bgl 
II 5.2 kb fragment were ligated to produce the plasmid pAdBglll. 
III.A.2. Construction of pAd5-CMV-lacZ 
A complete minigene containing the CMV enhancer and promoter, the lacZ 
gene, and the SV40 late gene polyadenylation signal was cloned into the Bglll site of 
AdBglll. The source of the minigene cassette was plasmid pCMV-0 (provided by 
Grant MacGregor, Baylor). A description and schematic of the construction of 
pCMV-0 have been provided by Dr. McGregor. The description is presented below 
verbatim and the schematic is provided in Panel B (Appendix F). 
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