M.J. Welsh and A.E. Smith, RAC Application 
2. Primary cultures of human airway epithelia grown on permeable supports. 
Procedures 
As described in Point B.2.b.(3), normal and CF airway epithelia were cultured on 
permeable filter supports and allowed to develop a low transepithelial conductance. They 
were then exposed to either the Ad2/CFTR-1 or the Ad2/BGal-l virus. 
Results 
The transepithelial conductance of normal and CF airway epithelial monolayers infected 
with Ad2/CFTR-1, Ad2/BGal-l, or no virus were indistinguishable. Figure 14A shows the 
time course of transepithelial conductance from 4 CF monolayers studied for 9 days after 
application of either Ad2/CFTR-1 (5 MOI) or vehicle contr ol; Fig. 14B shows mean data 
from 7 control CF epithelial monolayers and 10 Ad2/CFTR-1 -treated (5 MOI) monolayers 
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Figure 14. Effect of Ad2/CFTR-1 on transepithelial conductance. 
followed for 3 days after infection. The data indicate that there was no significant effect of 
virus on transepithelial conductance; transepithelial conductance usually begins to increase 
after two weeks in culture, because the monolayers have a limited lifetime. Moreover, 
infection with either of the two recombinant viruses did not alter the electrolyte transport 
properties (amiloride-sensitive Na + absorption and cAMP-stimulated chloride secretion). 
When the infected and uninfected monolayers were compared by light microscopy, they 
were indistinguishable; Fig. 15 shows an example. 
Conclusion 
These data indicate that infection of epithelial monolayers with the recombinant adenoviral 
vectors did not produce any adverse effects on tight junctions, cell morphology, or the 
capacity for transepithelial electrolyte transport. The lack of alteration in transepithelial 
conductance, which is directly related to the permeability of the epithelial barrier, indicates 
that there was no effect on the integrity of the epithelium. 
3. Hamster studies. 
Several studies involved the intratracheal instillation of the Ad2/BGal-l viral vector into 
Syrian hamsters. The hamster is a permissive host for serotype C adenovirus (143,144) and 
has been used as a model system. 
[900] 
Recombinant DNA Research, Volume 16 
