10. Urine sample for analysis and bacterial and viral culture. 
11. Measurement of transepithelial electric potential difference in the nasal 
epithelium. 
12. Nasal brushings and biopsies as described below. 
V.D. Clinical protocol 
1. Patients will be admitted to the GCRC and placed in isolation in a 
hospital room with standard respiratory precautions. A respiratory isolation sign 
will be placed outside the door, and hand washing will be practiced upon entering 
and leaving the room. Visitors must wear gloves, gown, and mask, as per usual in 
reverse isolation procedures at the UNC Hospitals. If the patient leaves the room, a 
mask will be required. 
2. Preparation of viral dose: The Ad5-CB-CFTR will be obtained as a 
purified and quality characterized virus from the University of Michigan Adenoviral 
Core Facility (J.M. Wilson, P.I.). The virus will arrive via overnight carrier. It will 
be opened and its contents directly placed in a syringe in Room 775 of Burnett- 
Womack. It will be instilled into the patient’s nostril in the patient’s room at the 
GCRC. 
3. For application of the virus, the patient will be placed in a lateral 
recumbent position such that the recombinant adenovirus can be applied to the 
inferior turbinate and the solution will move to the dependent area of the inferior 
meatus (beneath the inferior turbinate). Topical lidocaine (2%) will be applied to 
the nasal mucosa immediately before application of virus. Under direct vision, the 
recombinant virus will be applied to the surface of the inferior turbinate utilizing a 
syringe pump, with application of approximately 2 ml being superfused over a 
period of 30 min. 
4. For control studies, the patient will be repositioned into the opposite 
lateral recumbent position and vehicle (2 ml) perfused onto the contralateral nasal 
turbinate over 30 minutes. 
5. Three different doses of Ad5-CB-CFTR will be used in a dose escalating 
fashion. 
Patient 
Volume 
Titre 
Dose 
1-3 
2 ml 
< 
00 
o 
T— 1 
2 x log pfu 
4-6 
2 ml 
3 x 10 y 
10 11 
6 x 10^ pfu 
2 x 10^ pfu 
7-9 
2 ml 
Assuming that the surface area (8 cm 2 ) of the nasal mucosa to be studied has 
2 x 10 7 cells, a dose of 2 x 10^ pfu’s will correspond to a dose of ~ 10 pfu/cell; a 
dose of 6 x l(r pfus will correspond to a dose of 300 pfu / cell; and a dose of 2 x 
10^ 1 pfus will correspond to a dose of 10,000 pfus/cell. We will assess the results of 
each dose before escalating to the next dose. 
Recombinant DNA Research, Volume 17 
[491] 
