detection of CFTR. Analysis of fluid recovered three days after infusion revealed: 
a) In fluid recovered from the left upper lobe - lacZ expression in 5.2% of cells and 
CFTR expression in 0.3% of cells; and b) in fluid recovered from the right upper 
lobe - CFTR expression in 3.4% of cells and no cells expressing lacZ. 
Approximately lCr cells were recovered in the bronchial brushings, all of which 
have been placed in culture. Analysis of these cells has been restricted to SPQ assay 
for CFTR function. 
IV.B.6. Is recombinant adenovirus likely to be expressed in the germline? 
As shown in the data described above (see IV.B.2.), the intravenous 
administration of adenovirus in a quantity equivalent to that applied to the nasal 
surface did not lead to lacZ expression in the germline cells. Likewise, no evidence 
of germline expression was seen in male rabbits and monkeys (see IV.B.2.) receiving 
parenteral lacZ adenovirus. 
IV.B.7. Recombination 
Homologous recombination does occur within adenovirus serotypes. To 
minimize in vivo recombination, patients will be screened for immunity to Ad5 and 
administration of Ad5-CB-CFTR restricted to seropositive patients. 
IV.B.8. Malignancy 
Adenoviruses can cause tumors in newborn animals and can transform cells 
in tissue culture. The transforming function identified in tissue culture involves the 
E1A region. This region has been deleted in the proposed vector. No human tumor 
has been causally related to adenovirus (87-89). 
IV.B.9. Integration 
The data for Karlsson et al. (90,91) indicate that integration, if it does occur, 
is very low in cultured cells. No studies have been performed to test for integration 
in airway epithelia with a restricted life-span. 
IV.B.10. Immune response 
Adenoviral infections are associated with an immune response. This 
response is important for clearing adenoviral infections. Studies to determine the 
effects of this response of the feasibility of repetitive dosing of adenoviral vectors 
are underway. 
V. HUMAN PROTOCOL 
VA. Experimental Design of Human Protocol 
VA1. Introduction 
The overall goal of the protocol will be to evaluate the safety and biological 
efficacy of CFTR gene transfer using recombinant adenoviruses administered to the 
nasal epithelium of patients with cystic fibrosis. The general design is a study of 
ascending dose for safety and for biological efficacy, with patients serving as their 
own controls for the treated side of the nose, and with contralateral sham-control 
nasal studies. At the completion of the study, we would expect to know 1) whether 
this recombinant adenovirus transfers the normal CFTR cDNA to nasal epithelial 
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Recombinant DNA Research, Volume 17 
