Recombinant DNA Advisory Committee - 12/^4/92 
application of the adenovirus vector to their left nare. If no adverse reactions are 
observed 24 hours after this preliminary application, the patient will receive 20 ml of the 
adenovirus vector into the left main stem bronchus. The right side of the patient will not 
be treated so that it will function as a control. 
Following vector administration, the patient will remain in a negative pressure room in 
order to minimize aerosols. Daily cultures will be obtained of the patient's sputum, 
stools, urine, and nasopharynx until there is no longer evidence of vector excretion or 
secretion. In the unforseen circumstance that a patient requires intensive care unit 
(ICU) level of care, there are 3 negative pressure ICU rooms available. Patient care 
personnel will wear gowns, gloves, masks, and take the same precautions required for 
infectious viruses. Once the patient is no longer secreting or excreting virus, he/she will 
be removed from the negative pressure room. This period of isolation is estimated to be 
approximately 10 days. 
Patients will continue to undergo further testing following this period. These studies will 
include monitoring of specific parameters relating to the patient's pulmonary function. 
Individuals will undergo nasal mucosa and bronchial cell sampling by either scraping of 
the nasal mucosa or intermittent bronchial lavage. The cells obtained from these 
procedures will be assayed for: (1) DNA of the CFTR gene, (2) RNA resulting from the 
CFTR gene, (3) protein expressed as a result of CFTR gene expression, and (4) cell 
conductivity changes. In summary, the investigators will be looking for in vivo evidence 
of electrophysiological improvements and in vitro evidence of gene insertion and 
expression. 
Dr. Parkman discussed the proposed adenovirus vector. Retrovirus vectors, which have 
been traditionally used for gene therapy, require cell division in order to become 
effectively integrated. Since bronchial epithelial cells do not divide indefinitely, efficient 
transduction with retrovirus vectors would be almost impossible. For this reason, the 
investigators have developed an alternative vector based on a Type 5 adenovirus. 
Naturally occurring Type 5 adenovirus is capable of producing upper respiratory 
symptoms in humans. This vector has had all of the Ela and most of the Elb genes 
removed. Ela and Elb are the genes that are required for the regulation of 
transcription; although this vector has the capacity to infect and transduce non-dividing 
cells, it is incapable of replicating. The proposed vector has a relatively consistent point 
of integration; therefore, some of the concerns accompanying the use of retrovirus 
vectors (e.g., random integration and oncogenic potential) are minimized in this setting. 
The major concern regarding the use of this adenovirus vector is that Type 5 
adenoviruses exist naturally in our population. What will be the effect on a person 
whose cells have been transduced with this adenovirus? Could a wild-type infection 
Recombinant DNA Research, Volume 16 
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