M.J. Welsh and A.E. Smith, RAC Application 
samples from the 4 week animals also showed evidence of a dose-dependent increase in 
Ad2 antibody titer but at approximately a 100 times lower level than in serum. 
Conclusions 
The intratracheal administration of the described doses of Ad2/fiGal-l into hamsters does 
not appear to cause any persistent pulmonary or systemic adverse inflammatory events. 
Additionally, infectious virions were not detectable in either lung lavage or feces of the 1 
week animals nor in one month high dose lung tissue samples. 
c. Studies in progress 
A separate study, designed specifically to focus on detection of viral DNA in various 
organs to assess the spread of added virus is in progress. In this study, male and female 
Syrian hamsters have been given Ad2/8Gal-l (1.4 x 10 10 PFU) by intratracheal instillation. 
At 7 days post-instillation, various tissues, including lung, gut and gonads, were removed 
and analyzed by PCR for the presence of Ad2/BGal-l DNA. Under conditions where 
vector DNA was readily detected in treated lung samples, we have observed no confirmed 
PCR signal in liver, kidney, ovary and testes, nor in the lungs of sentinal hamsters. This 
experiment is being repeated with other species and other tissues, but results so far suggest 
that no transmission of vector DNA to other tissues, including the gonads, has occurred. 
An additional study, utilizing wild-type Ad2 is also in progress. The objectives of this 
study are twofold, to confirm the reported susceptibility of Syrian hamsters to wild-type 
adenovirus serotype C infection (91,144), and to provide positive control tissues for both 
immunofluorescence localization of adenoviral protein within various tissues and organs. 
Six male Syrian hamsters will be given intranasal doses of 1 x 10 7 PFU of wild-type Ad2 
virus. At 3, 5 and 7 days after virus administration, two animals will be sacrificed and 
various organs harvested for analysis. Lung and blood samples will be analyzed for the 
presence of infective virions. Other organs (as listed in the previous study) will be 
analyzed by immunofluorescence localization methods for the presence of viral proteins. 
To improve sample size, an additional study, similar in design to study one, but using larger 
numbers of animals, will begin shortly. We are also repeating the experiment in cotton 
rats, another host permissive for adenovirus replication. Both of these studies will use 
Ad2/CFTR-1. 
Conclusions 
Taken together, these hamster data suggest little or no systemic inflammatory response to 
the viral administration. There was a mild, transient, pulmonary inflammatory response 
which appeared to be associated with the intratracheal administration of the described doses 
of adenoviral vector in the Syrian Hamster. Serum antibody to adenovirus was detected in 
all treated animals. No evidence of vector replication was obtained. 
4. Primate studies. 
a. Ad2/BGal-l applied to the nasal epithelium. 
Procedure 
The procedure for two monkeys (A and B) was described above, point B.2.b.(3). The 
monkeys were completely evaluated on days 1, 4, 7, 14, 21, 28, 42, and 63. The monkeys 
were also observed daily for any abnormal behavior or physical signs. 
[904] 
Recombinant DNA Research, Volume 16 
