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 will be collected by standard venipuncture technique. Blood/serum analysis for a systemic 
response will include: complete blood count (including platelets and differential WBC), erythrocyte sedimentation rate, 
electrolytes (including Na, K, Cl, and HC0 3 ), and general chemistries (including total protein, albumin, Ca, P0 4 , 
glucose, uric acid, alkaline phosphatase, total bilirubin, AST, LDH, BUN, Cr, and amylase). PT and PTT will also be 
evaluated at the times indicated. The hematology assessment will require 5 ml of blood. The chemistry assessment 
will require 10 cc of blood. ^ 
Urinalysis for a systemic response will include: specific gravity, albumin, glucose, acetone, bile, pH, casts/HPF, 
RBC/HPF and WBC/HPF. 
, 
Bacterial Cultures , 
Purpose: To assess the extent of infection of Pseudomonas and other organisms (both aerobic and anaerobic) which 
may contribute to CF lung disease. 
Methods: Bacterial cultures for Pseudomonas ( aeruginosa and cepacia ), Staphylococcus aureus, and other organisms ■ n 
(both aerobic and anaerobic) will be conducted on samples from sputum in all study patients. > r 
Cytology with Cell Count 
Purpose: To determine the presence of an inflammatory response. 
Methods: Sputum samples will be collected from all study patients. The cells will be dissociated into 2 mL of 
phosphate buffered saline. The cell suspension will be kept on ice until further studies can be performed. Evidence of 
an inflammatory response will be assessed by cytological examination of cytospin preparations of the cells using Wright 
stain. The cell differential count will be determined for each specimen. Cell morphology and cytopathic effects will be 
evaluated using the PAP stain. 
Inflammatory Cytokines and Mediators 
Purpose: To determine the presence of an inflammatory response. 
I 
Methods: Sputum samples will be collected from all study patients. An evaluation of inflammatory cytokines and 
mediators including IL-1f3, IL-8 and TNF [24, 25] , agents associated with inflammatory airway disease in CF, will be 
performed by the Genzyme Corporation Immunology Department or the participating hospitals. The utility of measuring 
these cytokines in sputum samples from CF patients has been demonstrated. [26] 
Assessment for Evidence of any Viral Shedding of Ad2/CFTR-2 
Purpose: To determine the presence of Ad2/CFTR-2. 
j 
Methods: Nasal swab, sputum, urine and stool samples will be collected from all study patients. All samples will be 
cultured for Ad2/CFTR-2 on virus-permissive 293 cells. Based on our studies in animals, we expect that live virus could 
be present in the initial swabs but then would disappear with time. Our initial studies in humans suggest that 
Ad2/CFTR-1 could no longer be detected by culture one day after administration. 
If there is viral replication, we expect that after an initial decline, the titer of live virus in the nasal swab would increase. 
There will be a determination whether wild-type or recombinant virus is produced by culture on HeLa cells, by restriction j 
enzyme analysis, and by sequencing. 
j 
Assessment of Evidence of Antibody Development to Ad2/CFTR-2 
Purpose: To assess: 1 ) Whether the patient is seropositive for antibody to wild-type adenovirus prior to treatment with 
Ad2/CFTR-2; 2) The patient's antibody response to Ad2/CFTR-2. 
[558] 
Recombinant DNA Research, Volume 20 
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