Genzyme Corporation Ad2/CFTR-2 Gene Transfer Protocol: Single 
One Kendall Square, Cambridge, MA 02139 Aerosol Administration to the Lung of CF Patients 
Methods: Venous blood (10 ml) will be collected by standard venipuncture technique. Antibody to wild-type adenovirus 
will be determined prior to treatment with Ad2/CFTR-2.. Antibody titers to wild type adenovirus and Ad2/CFTR-2 will be 
determined by ELISA. Should the patient exhibit an antibody response to adenovirus, further analysis will be conducted 
to determine whether the antibodies are neutralizing antibodies by means of a cell based anti-viral assay. These tests 
have been developed and will be performed by the Genzyme Corporation Immunology Department. 
RS-PCR 
Purpose: To identify mRNA for CFTR and therefore provide biochemical evidence that the cDNA construct introduced 
to the cell has expressed the message for the protein synthesis of CFTR. 
Methods: RNA template-specific PCR (RS-PCR) is a modification of RT-PCR in which cDNAs containing a unique 5' 
nucleotide tag are synthesized from specific mRNAs, and are distinguished from contaminating DNA in a subsequent 
PCR by priming from the unique sequence. We are conducting this analysis of the mRNA for CFTR rather than CFTR 
itself because of the very limited amount of CFTR found in the cell. The majority of this protein is sequestered in the cell 
membrane where it forms a chloride channel. This study will be performed on cells obtained on Day 7 by brushing as a 
measure of safety and to ensure that no consistent compromise has occurred due to vector administration. 
Pulmonary Function Tests 
Purpose: To assess patient's pulmonary function. 
Methods: Patients will undergo either complete or limited pulmonary function testing as designated in the protocol. 
Complete pulmonary testing will include spirometry, lung volumes (both helium dilution and plethysmographic), single 
breath diffusing capacity and pulse oximetry testing. From spirometry, forced expiratory volume in one second (FEV-j : 
% of predicted) and forced vital capacity (FVC: % of predicted) will be obtained. Lung volumes, in particular the residual 
volume (RV: % of predicted) and total lung capacity (TLC: % of predicted) will be measured. The single breath diffusing 
capacity (DLCO: % of predicted) will also be obtained. Finally, pulse oximetry will be performed to measure the 
saturated oxygen levels (Sa0 2 ). For visits where limited pulmonary testing is required spirometry and pulse oximetry 
parameter data will be obtained. All tests will be performed using the standard procedures of the Pulmonary Function 
Laboratory at participating hospitals. 
Chest X-Ray 
Purpose: To assess: 1. The patient's pulmonary disease. 2. Radiological evidence of inflammation (including 
infiltrates) or an adverse effect. 
Methods: PA and lateral X-ray of the chest will be performed in the Department of Radiology at participating hospitals 
using their standard procedures. The chest X-rays will be evaluated for the radiological appearance of infiltrates and 
bleeding, other evidence of an inflammatory response or other evidence of an adverse effect. 
High Resolution Low Voltage Thin Section Computed Tomography of the Chest 
Purpose: To assess: 1. The patient's pulmonary disease. 2. Radiological evidence of inflammation (including 
infiltrates) or an adverse effect. 
Methods: High resolution, low voltage, thin section computed tomography of the chest (HRCT) will be performed in the 
Department of Radiology at participating hospitals. Each CT scan will be obtained with a 9800 scanner with the 
patient's lungs suspended in deep inspiration. Sections 1.5 mM thick will be obtained at 10 mM intervals and will be 
reconstructed with a bone algorithm. All patients will undergo low voltage imaging to reduce the overall radiation dose 
and minimize radiation to the thyroid gland. The patient's mediastinum and lungs will be imaged in a manner consistent 
with low voltage technique. The thin section HRCT will be evaluated for the radiological appearance of infiltrates and 
bleeding, other evidence of an inflammatory response or other evidence of an adverse effect. 
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