alveoli. However, by 4 weeks, this had resolved, with only few areas of 
very mild inf lamination in the alveoli. In contrast, animals receiving Ad5 
had inflammation, edema and epithelial damage, with mild to moderate in- 
flammation persisting at 4 weeks. Studies are ongoing to evaluate serum 
antibodies and shedding (pharyngeal, rectal). 
To determine if these responses to the replication deficient recombinant 
adenovirus were transient or permanent, in a limited, anecdotal evaluation 
(CR.007) of remaining animals from a prior study in which cotton rats 
received intratracheal Ad-CFTR 10 11 pfu/kg, the lungs of cotton rats admin- 
istered Ad-CFTR 16 weeks previously were evaluated by morphology. Consis- 
tent with the concept that the inflammatory response in cotton rats associ- 
ated with respiratory administrations of Ad-CFTR (n=3) does not persist, 
morphologic assessment demonstrated they were no different than the lungs 
of animals receiving intratracheal vehicle (n=2) or a naive animal from the 
same group (n=l) . Further evaluation of the parenchyma of these animals 
demonstrated no difference among the 3 groups, with no edema and/or changes 
in epithelial cells in either the airways or the alveoli. 
With the same batch of AdCFTR, 10 11 pfu/kg was administered intratracheally 
to 4 cotton rats and in parallel, compared to vehicle (n=4) and naive 
animals (n=2) . (CR.007A). As with intratracheal administration with AvlCFl 
(CRC.002), over 10 days, there were no deaths. The inflammatory response in 
the AdCFTR group was similar to that of several with AvlCFl (CRC.002). 
To determine if a replication deficient recombinant adenovirus has lethal 
effects when administered to the respiratory tract of cotton rats, increas- 
ing amounts of AvlCFl were administered by the nasal route to cotton rats 
in comparison to Ad5 (CRLD50.010) (Figure 4.4-A). Evaluation over a period 
of 52 days demonstrated no deaths in cotton rats receiving 10 6 pfu/animal 
(n=4) , 5X10 6 (n=4) , 10 7 (n=4) , 5X10 7 (n=4; except for 2 animals that died 
at 28d and 42d post administration following fights with other animals) , 
10 8 (n=4) , 5X10 9 (n=4) , 10 10 (n=4) and 5xl0 10 (n=4) . In contrast, there were 
deaths in animals receiving Ad5 in a dose response fashion, as has been 
observed by others (G. Prince, NIAID, personal communication). The data for 
the Ad5 animals by day 50 included: 10 7 pfu/animal (n=4, 0 deaths), 5X10 7 
(n=4, 0 deaths), 10 8 (n=4, 0 deaths), 5X10 8 (n=4, 0 deaths), 10 9 (n=4, 0 
deaths), 5X10 9 (n=4, 1 death day 6, 1 death day 11), 10 10 (n=4, 2 deaths 
day 4, 1 death day 7), and 4X10 10 (n=4, 2 deaths day 4, 1 death day 9, 1 
death day 11) . 
From these studies it can be concluded that there are inflammatory/immune 
responses to respiratory administration of a replication deficient, 
recombinant adenovirus with the design used in this protocol. This response 
is not lethal. The data suggest that the inflammation does not persist, and 
that no chronic changes are observed in the respiratory epithelium in re- 
sponse to the vector. 
To evaluate the recombinant adenovirus related inflammatory response ob- 
served in cotton rats in a context closer to humans , nonhuman primates were 
exposed via the large airways to AdRSV./9gal, a vector similar to Ad-CFTR 
except for the promoter (Rous sarcoma virus long terminal repeat) and the 
exogenous gene (E. coli lacZ coding for /9-galactosidase) (see Appendix 2 
Recombinant DNA Research, Volume 16 
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