M.J. Welsh and A.E. Smith, RAC Application 
chloride transport defect. For comparison, in a recent human trial using an E3 replacement 
adenovirus vector, 1.6 x 10 7 PFU were administered (93). 
4. Evaluation after virus application. 
After application of the virus, we estimate that the patient will be evaluated for 
approximately 14-16 days in the hospital. During the hospitalization, the patient will be 
confined to the hospital room. If it is required that they leave the room for specific 
procedures, they will wear a mask to cover the nose and mouth during such times out of the 
room. Strict respiratory, enteric, and body fluid isolation will be used. The patient will not 
be discharged from the hospital until two consecutive cultures for live virus from nasal 
swabs are negative. (Note that 7-8 days may be required to determine that a culture is 
negative.) Thus, the duration of hospitalization could potentially be longer than two weeks. 
During hospitalization, the patient's standard therapies for CF will continue. To maximize 
safety, results from each patient will be evaluated before proceeding to the next patient. 
Specific assessments performed on each patient will include the following (the timing of 
specific assessments is given in the protocol, Appendix 1): 
a. 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. Vital signs will be recorded. The area of application of the recombinant 
virus will be visually examined by endoscopy and compared to nontreated epithelium. We 
will look for evidence of exudate, inflammation, edema, or erythema. 
b. Blood/serum by venipuncture. 
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 the blood count and chemistry evaluations described above in the pretreatment 
evaluation. Antibody titers to the recombinant Ad2/CFTR-1 adenovirus will be determined 
by ELISA. 
c. Viral cultures of nasal and pharyngeal swabs, nasal brushing, blood buffy coat, 
urine, and stools. 
Purpose: To determine the presence of recombinant virus. 
Methods: All samples will be cultured on virus permissive 293 cells. We expect live virus 
to be present in the initial swabs but then to disappear with time. If there is viral 
replication, we expect that after an initial decline, the titer of live virus in the nasal swab 
would increase. We will determine whether wild-type or recombinant virus is produced by 
restriction enzyme analysis. 
Recombinant DNA Research, Volume 16 
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