Lung function (spirometry, He dilution, DLCO, body box, ABG) 
Sputum (volume, culture) 
Bronchoalveolar lavage (cell number, type, culture, volume) 
Nasal brush (cell number, type, culture) 
Bronchial brush (cell number, type, culture) 
The total maximum radiation exposure for roentgenographic and scintigraphic 
parameters for the first year of the protocol is approximately 4 rads. This 
amount is judged safe by NIH guidelines (less than 5 rads per year allowable). 
5. 5. 2. 2 Vector-related safety parameters 
The safety parameters directly related to the AdCFTR vector include (see 
Section 5.6 for specific times of evaluation): 
Anti -Ad antibodies (serum, lavage) 
Adenovirus culture (nasal brush, bronchial brush, pharynx, blood, 
rectal, urine) 
Adenovirus DNA - (nasal brush, bronchial brush, blood) 
5.5.3 Efficacy Parameters 
Evaluation of the efficacy of a recombinant adenovirus containing the human 
CFTR cDNA to treat the respiratory manifestations of cystic fibrosis will be 
based on biologic and clinical parameters. 
5. 5. 3.1 Biologic Efficacy Parameters 
The focus of these parameters is to demonstrate that the recombinant vector 
will compensate for the endogenous abnormal CFTR genes to provide normal CFTR 
gene-related expression to the airway epithelial cells. Most of these parame- 
ters will be evaluated in nasal and bronchial airway epithelial cells obtained 
by brushing the epithelium (see description of methods in sections 3, 4 and 
references Chu et al . , 1991; Trapnell et al . , 1991a). The epithelial cells 
will be recovered periodically during the baseline period (see section 5.6.2), 
the vehicle control period (section 5.6.3) and during the AdCFTR experimental 
treatment period (see section 5.6.4). The times of assessment of each parame- 
ter are detailed in section 5.6. The parameters to be evaluated (see sections 
3.1, 3.2 for details as to methodology), include: 
Expression of normal CFTR genotype at the mRNA level (sequence, PCR with 
specific probes) 
Level of total CFTR mRNA (quantitative PCR, Northern, in situ hybrid- 
ization analysis) 
Secretion of Cl" in response to intracellular elevation of cAMP ( 36 C1~ 
efflux, SPQ dye) 
Expression of CFTR protein ( immunohis tochemis try , immunoprecipitation and 
phosphorylation with kinase reaction, 35 S -methionine metabolic labeling 
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