Pulmonary function tests 
Pulse oximetry and arterial blood gas 
Electrocardiography (12-lead) 
PA and lateral chest x-ray 
Thin-cut CT scan 
Sputum culture with antibiotic sensitivities 
Shwachman-Kulczycki Score calculation 
Pretransfection treatment 
Beginning 2 weeks prior to transfection, the patient will begin an intensified treatment protocol 
to reduce respiratory infection and maximize overall condition. For the two weeks, the patient 
will receive two anti-Pseudomonal antibiotics to which their cultured organism is sensitive. 
Twice a day postural drainage and percussion will be performed. The patient will continue on 
the remainder of their chronic treatment regimen. This phase can be accomplished either as an 
inpatient or outpatient. During the subsequent studies, the patient will continue on their 
previously prescribed medical program. This includes continuation of any oral antibiotics, 
pancreatic enzymes, theophylline, and vitamin supplements. Aerosolized bronchodilators and 
antibiotics will be continued. 
Selection of segment for transfection 
The chest x-ray and thin cut CT scan will be used to select an anatomical pulmonary segment 
that: 
a) Has a degree of disease involvement average for that patient 
b ) Is in a location such that the patient can be positioned at bronchoscopy so that the 
segmental bronchus is gravitationally dependent 
Transfection procedure 
The patient will be prepared for bronchoscopy as indicated above. The bronchoscope will be 
introduced and advanced to the orifice of the segment chosen for the transfection. Secretions 
present within the segmental bronchus will be aspirated through the bronchoscope. 
Transepithelial electrical potential difference will be measured in the segment selected for 
transfection and in the segmental bronchus that is located in the same position in the opposite 
lung. A balloon catheter will be introduced through the bronchoscope channel and advanced one 
centimeter into the orifice of the lung segment to be transfected. The balloon will be inflated 
under direct vision until the orifice is minimally occluded. 50 ml of virus at a concentration of 
10 10 pfu/ml in normal saline at 37°C will be instilled through the balloon catheter. The 
catheter with balloon inflated will be held in place for 30 minutes, after which time any 
remaining fluid will be aspirated. The balloon will be deflated and the catheter and 
bronchoscope removed. 
In this initial study we propose to use a single dose of virus, 10 10 pfu/ml in a total volume of 
50 ml. This particular dose was selected based on our experience in human CF xenografts. We 
found that increasing the concentration of virus above 10 10 pfu/ml did not appreciably increase 
the efficiency of gene transfer. We elected not to try lower doses because of the very real 
possibility that the efficiency of gene transfer would be insignificant. In the absence of gene 
transfer little would be learned about safety and feasibility and the risks of 4 bronchoscopies 
would not seen acceptable.. 
Recombinant DNA Research, Volume 16 
[823] 
