M.J. Welsh and A.E. Smith, RAC Application 
Ad2-ORF6/PGK-CFTR 
Item 5 - Clinical Protocol 
Vector Application. 
Patients will be admitted to the Clinical Research Center at the University of Iowa. After vector 
administration, they will be placed in isolation in a hospital room for one day. They will be discharged 
from the hospital the day after Ad2-ORF6/PGK-CFTR administration. 
We will administer Ad2- ORF 6 /PG K-CF 1 K by directly applying it to the nasal epithelium over the course 
of 30 min. We will use the epithelium of the entire nasal cavity, which we estimate to be 20 to 50 cm 2 . 
On the first administration, the nasal epithelium that will receive Ad2-ORF6/PGK-CFTR will be chosen at 
random. We will administer saline to the opposite nasal epithelium. After administration, we will remove 
any excess fluid. The same nasal cavities will be treated with Ad2-ORF6/PGK-CFTR or saline on all 
subsequent administrations. 
Dose of Vector. 
Each participant will receive the same doses of Ad2-ORF6/PGK-CFTR: 
Administration Total IU 
1 2x107 
2 2x108 
3 2xl0 9 
4 lxlO 1 ® 
5 1x10*0 
Timing Of Repeat Administrations. 
We estimate, based on our previous studies in non-human primates and in humans with Ad2/CFTR-1, 
that the duration over which an adenovirus vector may correct the CF Cl" transport defect will be 
approximately 4 weeks, but could be substantially longer. The planned interval between vector 
administrations will be 4 weeks. However, the interval may be extended at the discretion of the 
investigators if the correction of the Cl" transport defect persists beyond 4 weeks or if unavoidable 
problems should arise. For Part A of this study which is focused on safety, precise timing between 
administrations will not be critical to the evaluation or interpretation of the data. However, the shortest 
duration between treatments under any circumstances will be 4 weeks. 
List Of Evaluation Procedures. 
History and physical examination . 
Purpose : To obtain evidence of patient discomfort, systemic responses, or inflammation. 
Methods : The patient will be questioned about local or systemic manifestations of inflammation. Specific 
attention will be placed on symptoms of nasal congestion. Vital signs will be recorded. The area of 
application of the recombinant virus will be visually examined by endoscopy and compared to nontreated 
epithelium. In many cases the area of administration will be photographed to provide documentation. We 
will look for evidence of exudate, inflammation, edema, or erythema. 
BlQQd/SgQim by ve ni puncture . 
Purpose : To assess the systemic response to virus application and the antibody response to recombinant 
virus. 
Methods : Venous blood will be collected by standard venipuncture technique. Analysis will include: 
complete blood count (including platelets and differential WBC), erythrocyte sedimentation rate, 
electrolytes (including Na, K, Cl, and HCO 3 ), and general chemistries (including total protein, albumin, 
Ca, PO 4 , glucose, uric acid, alkaline phosphatase, total bilirubin, AST, LDH, BUN, Cr, and amylase). 
PT and PTT will be evaluated at the times indicated. Antibody titers to adenovirus will be determined by 
ELISA and by determination of neutralizing antibodies. 
Viral cultures of nasal and pharyngeal swabs, urine, and stools . 
Purpose : To determine the presence of recombinant virus. 
Methods : All samples will be cultured on virus-permissive 293 cells. Based on our studies in animals, 
we expect that live virus could be present in the initial swabs but then would disappear with time. Our 
initial studies in humans (Appendix 7.8) suggest that Ad2/CFTR-1 could no longer be detected by culture 
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