Table I 
Organ Specific Expression of Human CFTR mRNA in Transgenic Mice: 
Detection by In Situ Hybridization Analysis of Individual Organs 1 2 
Organ Evaluated 2 
Promoter 
Lung 
Testes 
Ovarv 
Brain 
Uterus 
SDleen 
Kidnev 
SP-C 
+ + 
+ + 
- 
- 
- 
- 
- 
CC 10 
+ 
+ + 
+ 
+ 
+ 
+ 
fl-Actin 
+ + 
- 
- 
- 
- 
1 Mice carrying transgenes consisting of the human CFTR cDNA driven by various 
promoters as indicated 
2 Expression in a particular organ is indicated by a " + " or " + + " (more abundant 
expression). Absence of expression in an organ is indicated by 
transgenic animals to date (Figure 25). 
In summary, evidence to date derived from transgenic mice in vivo, cotton rats infected with 
adeno-CFTR, human respiratory epithelial cells grown on tracheal grafts transfected mammalian 
cells in vitro, all support the safety of expressing the CFTR mRNA at varying levels in a variety 
of distinct cell types. 
4.3 In Vivo Use of a Replication Deficient, Recombinant Adenoviral Vector to transduce 
Epithelial Cells of the Human Respiratory Tract: Safety concerns in this area are divided into 
questions regarding 1) toxicity, 2) the potential for vector-induced immune responses, 
inflammation and tissue damaged related to replication of the vector, and 3) alterations in the cell 
physiology and functions other than those related to CFTR in transduced respiratory epithelial 
cells and 4) malignancy. 
4.3.1 Host Response Following Respiratory Administration of AE1, Recombinant 
Adenovirus: Will the vector express viral genes? If so, will viral proteins expressed within the 
epithelium or which are part of the initial administration of vector result in a local humoral or 
cellular immune response? Will a systemic immune response occur following respiratory 
administration of AvlCF2? Will the patients become immune to the adenovirus vector? What 
is the relevance, if any, of an E3 vector deletion to immune response in AE1 vectors? These 
related questions will be addressed below. 
It is anticipated that the vector may express some viral genes at a very low level. This is 
based on results observed in vitro in cultured epithelial cell lines infected with similar vectors 
(Ad-CFTR, AvlCFl (Crystal et.al., RAC proposal, 1992); Ad2/CFTR-1 (Welsh et.al., RAC 
proposal, 1992); and Ad.CB-CFTR (Wilson et.al., RAC proposal, 1992). In culture, HeLa cells 
infected with control adenovirus rapidly produce large amounts of hexon protein, a major capsid 
protein. However, AE1 vector infected cells produced only minimal amounts of this protein. 
Similar results have been observed in AvlCF2 infected Hela cells (not shown) were levels of 
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Recombinant DNA Research, Volume 17 
