M.J. Welsh and A.E. Smith, RAC Application 
gene and second, deletion of DNA sequences to enable the resulting recombinant to 
become small enough to package its DNA. The most likely sequences to be deleted, 
compatible with retention of viral replication, would be some or all of the CFTR DNA. 
Growth of such a recombinant would probably be self-limiting and would be unlikely to 
lead to synthesis of CFTR fragments with any significant biological activity. 
Our animal and cultured cell studies, some using doses greatly in excess of the human dose, 
have revealed no evidence for Ad2/CFTR-1 replication. Moreover, we found no evidence 
for Ad2/6Gal-l replication despite its smaller size and more ready growth in 293 cells. 
c. Effects of wild-type adenovirus 2 infection. 
Potential effects of wild-type Ad2 include symptoms of upper respiratory infection. 
Pharyngitis and conjunctivitis are known to be associated with adenovirus. Strains of 
adenovirus other than Ad2 have been known to cause pneumonia, transient diarrhea and 
gastroenteritis. Rare problems associated with adenoviral strains include cystitis and 
keratoconjunctivitis. Such problems could be hazardous in immunocompromised hosts. 
Risk is minimized in the protocol by the use of patients who have only mild to moderate 
disease and who have evidence of preexisting immunity to adenovirus. The possible role 
of adenovirus in oncogenesis is discussed in Section 4.4; there is no evidence to suggest a 
role for adenovirus in human tumors. 
d. Risks associated with the study procedures. 
i. Measurement of the transepithelial electrical potential difference across the nasal 
epithelium will not produce significant discomfort; it has generally been well tolerated in 
the past. The drugs applied topically to the nasal epithelium during the course of the study 
have no significant local or systemic effects. 
ii. Nasal and pharyngeal swabs can produce mild discomfort when they are taken, but there 
is no serious or long term risk. They are standard practices in clinical outpatient medicine. 
iii. Brushing the nasal mucosa could produce discomfort that might persist for several 
hours. It could also produce some minor bleeding (a few ml at most). There are no major 
or long term risks. 
iv. The process of application of the recombinant virus to the nasal mucosa should have no 
significant risk. There could be minor discomfort or a sense of nasal obstruction. There 
may also be discomfort from having to remain still during the procedure. These 
discomforts should be minimized by local anesthesia with topical 2 % lidocaine to the nasal 
mucosa and by administration of Midazolam 1-2 mg. IM. shortly before application. The 
major risks of lidocaine and Midazolam would be hypersensitivity which is very rare. The 
Midazolam could theoretically produce some respiratory depression; however the dose we 
will use should have no significant effects, particularly in these subjects who will only have 
mild to moderately severe disease. 
v. Biopsy of the nasal epithelium will produce mild to moderate pain and discomfort. Pain 
and discomfort during the procedure will be minimized by use of local anesthesia. 
Subsequent use of analgesics will be allowed if needed. A small risk of bleeding or 
subsequent scar formation after the biopsy is possible as is expected for such a routine 
procedure. There is also a risk of infection. We will treat pain with analgesics and observe 
the area for bleeding or infection; if either should require it, we will administer appropriate 
treatment. 
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Recombinant DNA Research, Volume 16 
