M.J. Welsh and A.E. Smith, RAC Application 
We found that a relatively low 
multiplicity of infection with 
Ad2/CFTR-1 was able to generate 
apparently wild-type transepithelial 
chloride secretion; in fact, we detected 
significant chloride secretion at an 
MOI of 0.1. Secretion occurred at low 
multiplicities even though the 
promoter directing expression of 
CFTR is the E la promoter which is not 
regarded as particularly strong. This 
result is consistent with those reported 
above in Section 3.3, suggesting that 
only modest expression of CFTR is 
required for phenotypic correction. 
We believe that this represents the 
most significant data we have 
suggesting the efficacy of the 
Ad2/CFTR-1 vector. It uses human 
CF cells; it examines them under 
conditions that most closely resemble 
an in vivo epithelial monolayer; it 
measures the function of CFTR; and it 
shows that only small amounts of virus 
may be effective. 
3. Primate studies. 
We have done three types of studies to 
assess the efficacy of the adenovirus 
constructs in the airway epithelia of 
Rhesus monkeys. 
a. Ad2/BGal-l applied to the nasal 
epithelium. 
Procedure 
For this study we have used 2 monkeys 
(A and B). As of late September, they 
have been followed for 8 weeks after 
virus administration. 
Because primates have normal CFTR 
function, we elected to assess the 
ability of recombinant adenovirus to 
transfer the reporter gene for B-galactosidase by using the Ad2/BGal-l virus. We estimated 
the epithelial cell density in the nasal cavity of the Rhesus monkey to be 2 x 10^ cells/cm^ 
(based on an average nasal epithelial cell diameter of 7 pm, ref. 140) and the surface area to 
be 25-50 cm^ (141). Thus, we estimated that there are about 5 x 10^ cells in the nasal 
epithelium of Rhesus monkey. Because these studies were also focused on safety, we used 
higher viral doses. 
Figure 10. Immmunocytochemical 
staining of CFTR in CF airway epithelia 
infected with Ad2/CFTR-1. 
Recombinant DNA Research, Volume 16 
[891] 
