shedding, immunologic reactions, or other potential adverse effects from AAV-CFTR 
vectors delivered to the airway surface via a flexible fiberoptic bronchoscope. 
In order to determine the biological activity and safety of AAV-CFTR vector administration 
in primates a study of AAV-CFTR vector administration was performed in rhesus 
macaques. Doses of vehicle, 5x10^, 1x10^, or 1x10^ 1 particles of AAV-CFTR were 
administered to the RLL through a fiberoptic bronchoscope. Animals were sacrificed at 10 
days, 21 days, or 90 days after vector instillation. 
Vector DNA was detectable in cells from the RLL of the vector-treated animals, but not in 
the control animals. The presence and expression of AAV-CFTR vector DNA was also 
detected in the liver of the 10^ dose animal, and in the trachea. There were no significant 
abnormalities noted in any of the safety assessments, although there was a mild prominence 
of the bronchus-associated lymphoid tissue (BALT) in the 10^ dose monkey. The 
functional studies of airway function (PFT) and gas exchange (arterial blood gases, chest 
radiographs) were entirely normal. Although spread to the liver was observed by in situ 
PCR it was localized to one small area (<10% of the total area of one cross section), and 
there was no spread to the gonads, even at this very high dose, and no evidence of 
neoplastic changes. Taken together these studies indicate that AAV-CFTR vector 
administration is safe in primates at doses ranging from 5x10^ to 1x10^ particles 
administered to a single lung lobe and at time points ranging from 10 days to 90 days after 
vector instillation. 
One additional safety issue which has been studied in rhesus macaques is that of the 
potential for spread of recombinant AAV-CFTR from vector-treated individuals who might 
subsequently be infected with wild-type AAV2 and adenovirus. In cultured cells 
previously transduced with AAV-CFTR recombinants, infectious vector can be "rescued" 
when they are infected with wild-type AAV and adenovirus. In order to determine whether 
this phenomenon would occur in the complex context of an in vivo infection, we studied 
two rhesus monkeys, both of whom were infected with 1x10*0 particles of AAV-CFTR to 
the RLL. Ten weeks later these animals were infected intranasally with an adenovirus type 
2 host-range mutant (Ad2HR405), which is known to possess AAV2 helper function and 
to be able to infect monkey cells in culture, and with wild-type AAV2. Viral cultures were 
performed prior to infections and at 3 days, 7 days, 14 days, and 21 days from 
bronchoalveolar lavage (BAL) fluid and nasal washings. The results of these studies were 
as follows: Ad2HR405 infection was clearly established since adenovirus replication was 
detectable in both BAL and nasal wash cultures from the 3 day and 7 day samples, while 
none was seen in the pre-treatment fluids. AAV2 replication was also established in one of 
the two monkeys at 3 days, but ONLY in the nose. No recombinant AAV-CFTR was 
detectable in any fluid at any time point. These data indicate that while rhesus monkeys can 
have experimental adenovirus and AAV infections established, the AAV infection remains 
localized to the upper respiratory tract, near its normal portal of entry, and does not appear 
to spread to the lower tract. Since AAV replication does not occur in the lower tract where 
vector is localized, there appears to be little opportunity for in vivo rescue to occur. 
4.0 STUDY DESIGN 
This will be a single center, phase I, open-label, non-randomized, dose escalation study in 
1 6 patients with cystic fibrosis and mild lung disease. Each individual will receive a single 
dose of AAV-CFTR vector administered to the nasal epithelium alone or to the nasal 
epithelium and the right lower lobe (RLL) of the lung. 
Recombinant DNA Research, Volume 20 
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