M.J. Welsh and A.E. Smith, RAC Application 
not detected in stool samples taken for over one month, suggesting that ingestion of virus 
followed by replication in the gut does not occur. 
b. Ad2/CFTR-1 applied to the nasal epithelium. 
Procedure 
The procedure for three monkeys (C, D, and E) was described above. Point B.2.b.(3). The 
monkeys were completely evaluated on days 1, 3, 6, 13, 20, and 27. The monkeys were 
also observed daily for any abnormal behavior or physical signs. 
Results 
Examination. The three monkeys tolerated the application procedure well. Visual 
inspection of the nasal epithelium revealed no inflammatory response. Daily examination 
revealed no evidence of coryza or conjunctivitis. There was no cough, sneezing, or 
diarrhea. Slight erythema was observed in all three monkeys in both the control (left) and 
virus (right) nostril on the first day after infection and then disappeared. Appetites and 
weights were not affected by virus administration. Monkey E had a poor appetite requiring 
special diet before the infection, but it improved throughout the experiment. Physical 
examination revealed no evidence of fever, lymphadenopathy, tachypnea or tachycardia at 
any time. 
Blood counts and serology. One of the monkeys had a relative blood leukocytosis on day 
6 post infection. The WBC increased to 8,000 from a preinfection count of 3,400. No 
abnormalities were found in the hematology of the other two monkeys . Serum ESR, 
electrolytes, transaminases, BUN and creatinine were normal throughout. 
Cytology and Histology of the nasal epithelium. We assessed epithelial inflammation by 
cytological examination of Wright-stained cells obtained from brushings of the nasal 
mucosa. Wright stains of cells from the nasal mucosa were evaluated at each of the time 
points. Monkey C had a slight increase in inflammatory cells in both control and infected- 
nostrils on day 1; on subsequent days the number of inflammatory cells (lymphocytes, 
neutrophils, eosinophils) was less than 5%. Monkey D had less than 5% inflammatory 
cells throughout. Monkey E had an increased percentage of eosinophils in both nostrils at 
all the time points. 
Presence of virus on the airway epithelium. We tested for the presence of virus in the 
supernatant of the cell suspension from swabs and brushes from each nostril and the 
pharynx. Cytopathic changes were monitored for 5 days and then the 293 cells were 
stained with an FITC-labeled anti-adenovirus monoclonal antibody (Chemicon). Live virus 
was detected on day 1 and 3, but not on day 7 or thereafter. 
Stool culture. Stools were collected every week and cultured for virus; all were negative. 
Adenovirus antibody titers. All three monkeys had an increase in adenovirus titers and 
adenovirus 2 antibody titers after infection. 
Conclusions 
None of the monkeys developed any signs associated with wild-type adenoviral infection. 
Significant eosinophilia was observed from nasal brushings of one of the monkeys; this 
was not associated with blood eosinophilia or with clinical signs of allergic rhinitis. As we 
indicate below, this was also observed in the bronchial brushings of some of the monkeys 
before virus administration and it was found in the sentinal monkey. Monkeys that had 
eosinophilia on any respiratory surface were monkeys E, F, G, and H. These four monkeys 
were recently transferred from San Francisco to Iowa; monkeys A and B have lived in Iowa 
[906] 
Recombinant DNA Research, Volume 16 
