Novel adenovirus gene therapy vehicle and cell line

Abstract

A novel adenovirus E1/E4 expressing packaging cell line is provided, which permits the generation of recombinant adenoviruses deleted in both gene regions. The E1/E4 deleted recombinant adenovirus is capable of expressing a selected transgene product in cells in vivo or in vitro. This recombinant virus is useful in the treatment of genetic disorders.

What is claimed is:  
     
         1 . A packaging cell line which expresses an adenovirus E1 gene and an adenovirus E4 gene or a functional fragment of said E1 or E4 genes.  
     
     
         2 . A stable packaging cell which comprises: 
 (a) a first adenovirus DNA under the regulatory control of a first promoter, wherein said first adenovirus DNA encodes an adenovirus E1 protein; and    (b) a second adenovirus DNA under the regulatory control of an inducible promoter, wherein said second adenovirus DNA encodes an adenovirus E4 ORF6 protein,    wherein said packaging cell contains no other adenovirus E4 ORF DNA; and    whereby, in the presence of an inducing agent, said E1 and E4ORF6 proteins are expressed in amounts sufficient to permit infection by, and recovery of, an adenovirus having functional deletions in its E1 and E4 genes.    
     
     
         3 . The cell of  claim 2 , which is a human cell.  
     
     
         4 . The cell of  claim 2  in which the inducing agent is a metal.  
     
     
         5 . The cell of  claim 2  in which the metal is zinc.  
     
     
         6 . The cell of  claim 2  in which the inducing agent is a glucocorticoid.  
     
     
         7 . The cell of  claim 2  in which the inducing agent is dexamethasone.  
     
     
         8 . The cell of  claim 2  in which the first adenovirus DNA and the second adenovirus DNA are within the same nucleic acid molecule.  
     
     
         9 . The cell of  claim 2  in which the nucleic acid expressing the adenovirus E4ORF6 protein is pMMTVE4ORF6.  
     
     
         10 . The cell of  claim 2  in which the nucleic acid expressing the adenovirus E4ORF6 protein is pMTE4ORF6.  
     
     
         11 . The cell of  claim 2  which is 293 (MT-ORF6).  
     
     
         12 . The cell of  claim 2  which is HeLa (MT-ORF6).