V.E. Evaluation of adenoviral effect 
Introduction . After the virus is instilled into the nasal epithelium, we 
estimate that the patient will be studied in the GCRC for 14 to 21 days. The patient 
will be confined to the GCRC room except as required for tests out of the room. 
During such times, patients will wear a mask to cover the nose and mouth, and strict 
respiratory and body fluid isolation will be accomplished. The patient will not be 
discharged from the hospital until two consecutive viral cultures are negative. The 
usual therapies for CF will continue through the study protocol. 
V.E.l. To assess for safety: 
V.E.l.a. Inflammation 
V.E.l.a.(i) Patients will be monitored daily for symptoms of local 
or systemic inflammation. The site of application of the recombinant virus and 
control nostril will be compared by nasal endoscopy. Evidence of inflammation will 
be recorded. 
V.E.l.a.(ii) The nasal epithelium will be biopsied for assessment 
of inflammatory infiltrate and for evaluation of cytopathic effects. This procedure 
will be performed by our ENT colleague, Dr. Pillsbury. The patients will be 
anesthetized with solution of topical anesthetic and epinephrine placed under direct 
vision. The region of the biopsy will be then infiltrated with anesthetic and 
epinephrine and 0.5 cm^ of nasal mucosa will be resected by biopsy forceps with a 
cotton pledget placed at the biopsy site to effect hemostasis. The biopsy will be 
assessed for integrity of the epithelium and for evidence of inflammatory changes by 
light microscopy. Electron microscopy will also be performed to assess for 
cytopathology. 
V.E.l.a.(iii) Blood/serum will be monitored serially for 
assessment of humoral antibody response to the recombinant virus and for any 
systemic response after the viral application. Blood and urine will be collected, and 
antibody titers to Ad5-CB-CFTR will be assessed by ELISA. Nasal lavages will be 
assessed for evidence of IgA antibody titers to Ad5-CB-CFTR. Blood and serum 
analyses will include CBC with white count and Chem 18 performed in UNC 
Hospital labs. The blood buffy coat, urine and stool will be cultured in a plaque- 
forming assay on 293 cells to assess for virus. 
V.E.l.b. Shedding 
Sequential swabs of the nose and the pharyngeal mucosa (and nasal lavages) 
and urine and stool samples will be undertaken to assess for viral shedding by 
culture and to assess inflammatory response by histology. Live recombinant virus 
will be tested for by plaque-forming assay on the 293 cell line. The virus is likely to 
be present for the first several days but disappear after a week. Evidence of 
inflammatory response will be assessed by counting inflammatory cells in the nasal 
wash specimen. 
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Recombinant DNA Research, Volume 17 
