3.1.1 Expression of CFTR mRNA 
3.1.2 Expression of CFTR protein 
3.1.3 Functional "complementation" of the cystic fibrosis phenotype 
3.1.4 Can the vector survive in the airway epithelial lining fluid of 
individuals with cystic fibrosis? 
3 . 2 In vivo evidence 
3.2.1 Expression in vivo in cotton rats 
3.2.2 Expression in vivo in non-human primates 
4. Safety considerations 
4.1 Will the vector replicate in human epithelial cells? 
4.2 Does the vector express viral genes in addition to the CFTR cDNA? 
4.3 Is respiratory infection with the vector associated with transfer of 
the exogenous gene to the germ line? 
4.4 Is respiratory administration of the vector associated with damage to 
the lung? 
4.5 Are anti-vector antibodies elicited by respiratory administration of 
the vector? 
4.6 Does repeat administration of the vector pose a risk? 
4.7 Is the vector shed following respiratory administration of the 
vector? 
4.8 Are there risks of having some of E3 deleted from the vector? 
4.9 Can recombination or complementation of the vector occur in vivo 
following respiratory administration of the vector, and if so, does 
this pose a risk to the patient and/or environment? 
4.10 Is there a risk of malignancy associated with the use of the vector? 
4.11 Does the genome of the vector integrate into the genome of the target 
cells? 
4.12 Does the expression of the CFTR cDNA need to be regulated? 
5. Human Protocol 
5.1 General design 
5.2 Constraints that dictate design of the protocol 
5.3 Baseline safety data from individuals with cystic fibrosis relevant 
to administration of a replication deficient recombinant adenovirus 
5.3.1 Anti-vector antibodies in blood and lungs of individuals with 
cystic fibrosis 
5.3.2 Infectious viruses in the respiratory tract of individuals with 
cystic fibrosis 
5.3.3 Baseline inflammation in nasal epithelium, bronchial epithelium 
and epithelial lining fluid of individuals with cystic fibrosis 
5.3.4 DNA containing Ela sequences in nasal epithelial cells, 
bronchial epithelial cells, lung inflammatory cells, blood 
lymphocytes and blood neutrophils of individuals with cystic 
fibrosis 
5.4 Patient eligibility and selection 
Recombinant DNA Research, Volume 16 
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