8 . 
HIV infected individuals. 
9. Chronic active hepatitis B surface antigenemia or unstable hepatitis C. 
5.2.4 Methods 
Study Design: Patients will be admitted to Children’s Hospital Medical Center in a negative 
pressure isolation room on respiratory isolation procedures. See Table III for a summary of 
the clinical protocol. 
Day Minus 30 to Minus 18: Eligible cystic fibrosis patients will be seen in the clinical research 
center for pre-study evaluations. Fully informed consent will be obtained. The patients will 
have a baseline history and physical examination. They will also have baseline studies of EKG, 
chest x-ray, oxygen saturation in room air, pulmonary function tests, nasal brushings and a 
nasal biopsy, bronchoscopy for bronchoalveolar lavage, cells by brushing and biopsy, 
adenovirus culture for wild adenovirus infection, PCR for wild adenovirus infection, adenovirus 
serology, complete blood count, differential cell count and platelet count, prothrombin time and 
partial thromboplastin time, serum chemistry, urinalysis, sputum culture, genetic analysis for 
aF 508, sweat chloride test, an ELISA test for HIV, and (if appropriate) a pregnancy test. 
Presence of active adenoviral infection defined by culture or PCR will exclude patients from 
further study. The BAL fluid will be used for measurement of cellular composition, neutrophil 
elastase, and interleukin-8 concentrations. These are measurements of airway inflammation. 
2- week Cleanout: From days minus 18 to minus 4, the subjects in the study will receive 2 
weeks of intravenous antibiotics with the combination of a /3-lactam antibiotic and an 
aminoglycoside based on the results of prior and current respiratory cultures according to the 
standard divisional protocol. This will be combined with chest percussion and postural 
drainage, aerosol therapy, and optimization of nutrition. Optimal methods for "cleanout" 
include use of DNase (if licenced) will be utilized during this period. 
Day -3: History and physical examination will be repeated. Baseline chest x-ray, perfusion 
scan, CT scan of the chest, mucociliary clearance scan, oxygen saturation, and pulmonary 
function tests will be obtained. Nasal potential difference measurements will be made (see 
below), to document chloride permeability defect and feasibility of obtaining measurements in 
each patient. Measurements will be made at 5 identified sites measured along the inferior 
turbinates of each nostril from the anterior tip of the turbinate. Pharyngeal swabs and stool 
will be sent for culture for wild type adenovirus, PCR will be performed for wild type 
adenovirus, and adenovirus serology will be obtained. Complete blood count and differential, 
PT, PTT, serum chemistry, and urinalysis will be repeated. 
Day -2: History and physical examination will be repeated. Subjects will receive nasal 
treatment with the AvlCF2 vector. The nose will be anesthetized with 2% topical lidocaine 
and then examined by endoscopy. To facilitate patient cooperation, subjects may receive mild 
sedation with intravenous midazolam. 
The adenovirus-CFTR construct will be applied to an area of approximately 1 square cm 
in a defined region of the turbinates in a single nostril. The virus will be suspended in a 
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