M.J. Welsh and A.E. Smith, RAC Application 
h. Measurement of transepithelial electrical potential difference across the nasal 
epithelium. 
i. Pulmonary function tests including spirometry, lung volumes, and diffusing capacity. 
j. Arterial blood gases. 
k. Chest X-ray: P-A and lateral. 
l. Computerized tomography of the sinuses (at enrollment into the study and at the 
completion of the study only, unless indicated by a change in clinical status). 
3. Virus Application. 
Patients will be admitted to the Clinical Research Center at the University of Iowa on the 
day of virus administration. They will be placed in isolation in a hospital room with 
complete isolation precautions (see Appendix 4). 
Endoscopic examination of the nasal mucosa will be used to identify the optimal treatment 
area and to define well recognized landmarks. We will use the area along the inferior 
surface of the inferior turbinate. The area will be prepared by gentle rinsing with saline and 
suction. 
To minimize sneezing, discomfort, and movement during application of virus, patients will 
receive 2% topical lidocaine to the nasal mucosa immediately before application. As mild 
sedation and to prevent movement during the application, they will also receive Midazolam 
1-2 mg. IM. shortly before application. 
The recombinant virus will be applied to a defined area of nasal mucosa using an 
application area of approximately 0.5 cm 2 in each nares. We will use a plexiglass 
applicator that allows us to apply the virus to a defined area and then wash it away; in 
preliminary experiments we have tested the applicator in normal humans using methylene 
blue as a tracer. The virus will remain in contact with the nasal epithelium for 30 min. 
Then the mucosa will be gently washed to remove unattached virus. 
Different doses of the virus/gene construct will be used. 
PATIENT TOTAL DOSE* 
PFU 
Pg 
1 
2xl0 6 PFU 
0.25 
2 
2xl0 7 PFU 
2.5 
3 
5xl0 7 PFU 
6.25 
^Preparation and assay of production lots of virus is described under B.l.b.(l)(b). We will 
apply the virus in a volume of 50 to 200 p.1 to an area of approximately 1 cm 2 of nasal 
mucosa. Assuming that 1 cm 2 of nasal mucosa has about 2xl0 6 cells, a dose of 2xl0 6 PFU 
corresponds to a dose of approximately 1 PFU/cell. Based on our experiments with human 
airway epithelia, we expect that 10 PFU/cell will provide substantial correction of the CF 
[910] 
Recombinant DNA Research, Volume 16 
