Infectious papillomavirus pseudoviral particles

Abstract

The invention provides an infectious papillomavirus pseudoviral particle useful in gene transfer comprising: (a) a papillomavirus vector DNA which comprises an E2 binding site and an expression cassette comprising a gene and a sequence controlling expression of said gene; and (b) a papillomavirus capsid which comprises L1 and L2 structural proteins, such that said capsid encapsidates said vector DNA, wherein said gene is derived from a first biological species and said L1 structural protein is derived from a second biological species and said first biological species is different from said second biological species.

What is claimed is:  
     
         1 . An infectious papillomavirus pseudoviral particle comprising: 
 (a) a papillomavirus vector DNA which comprises an E2 binding site and an expression cassette comprising a gene and a sequence controlling expression of said gene; and    (b) a papillomavirus capsid which comprises L1 and L2 structural proteins, such that said capsid encapsidates said vector DNA.    
     
     
         2 . The infectious papillomavirus pseudoviral particle of  claim 1 , wherein each of said L1 and L2 structural proteins is derived from a human papillomavirus.  
     
     
         3 . The infectious papillomavirus pseudoviral particle of  claim 1 , wherein said gene is a human gene.  
     
     
         4 . A method of making infectious papillomavirus pseudoviral particles comprising: 
 (a) providing a cell line which expresses papillomavirus E2 DNA binding protein and L1 and L2 structural proteins;    (b) transforming said cell line with a papillomavirus vector DNA which comprises an E2 binding site and an expression cassette comprising a gene and a sequence controlling expression of said gene, wherein said papillomavirus E2 binding site is a cognate binding site of said E2 DNA binding protein;    (c) providing conditions for the encapsidation of said vector DNA by a capsid which comprises said L1 and L2 structural proteins to generate said particles; and    (d) harvesting said particles.    
     
     
         5 . The method of  claim 4 , wherein said cell line is a mammalian cell line, an insect cell line, or a yeast cell line.  
     
     
         6 . A cell line comprising the infectious papillomavirus pseudoviral particle of  claim 1 .  
     
     
         7 . Infectious papillomavirus pseudoviral particles made by the method of  claim 4 .  
     
     
         8 . A method of transferring a gene into a cultured mammalian cell comprising: 
 (a) providing the infectious papillomavirus pseudoviral particle of  claim 1;  and    (b) infecting a cultured mammalian cell with said particle such that said cultured mammalian cell is transformed with said gene.    
     
     
         9 . A method of screening for infectious papillomavirus pseudoviral particles comprising administering the infectious papillomavirus pseudoviral particles of  claim 7  as test particles to cultured non-infected mammalian cells and scoring for infectivity.  
     
     
         10 . A composition comprising the infectious papillomavirus pseudoviral particle of  claim 1 , wherein said gene encodes a product capable of having a therapeutic effect when administered in a therapeutically effective amount to a host subject in need thereof.  
     
     
         11 . A composition comprising the infectious papillomavirus pseudoviral particle of  claim 1 , wherein said gene encodes a product capable of having an immunogenic effect when administered in an immunogenically effective amount to a host subject in need thereof.  
     
     
         12 . An infectious papillomavirus pseudoviral particle for use as a medicament upon infecting cells of a human in vivo, wherein said particle comprises: 
 (a) a papillomavirus vector DNA which comprises an E2 binding site and an expression cassette comprising a gene and a sequence controlling expression of said gene, wherein said gene encodes a therapeutic protein and said cells express a therapeutically effective amount thereof; and    (b) a papillomavirus capsid which comprises L1 and L2 structural proteins, such that said capsid encapsidates said vector DNA.    
     
     
         13 . The method of  claim 12 , wherein said cells are epithelial cells and said therapeutic protein has a systemic effect.  
     
     
         14 . The method of  claim 12 , wherein said cells are epithelial cells and said therapeutic protein has a local effect on said epithelial cells.  
     
     
         15 . The method of  claim 13 , wherein said therapeutic protein is Factor IX and the expression of said therapeutic protein results in treatment of hemophilia.  
     
     
         16 . The method of  claim 14 , wherein said therapeutic protein is herpes simplex virus thymidine kinase and the expression of said therapeutic protein results in treatment of skin cancer.  
     
     
         17 . An infectious papillomavirus pseudoviral particle for use as a vaccine upon infecting cells of a human in vivo, wherein said particle comprises: 
 (a) a papillomavirus vector DNA which comprises an E2 binding site and an expression cassette comprising a gene and a sequence controlling expression of said gene, wherein said gene encodes an immunogenic protein and said cells express an immunogenically effective amount thereof; and    (b) a papillomavirus capsid which comprises L1 and L2 structural proteins, such that said capsid encapsidates said vector DNA.    
     
     
         18 . A second infectious papillomavirus pseudoviral particle, which differs from a first infectious papillomavirus pseudoviral particle, each particle for use as a vaccine upon infecting cells of a human in vivo, wherein each particle comprises: 
 (a) a papillomavirus vector DNA which comprises an E2 binding site and an expression cassette comprising a gene and a sequence controlling expression of said gene, wherein said gene encodes an immunogenic protein and said cells express an immunogenically effective amount thereof; and    (b) a papillomavirus capsid which comprises L1 and L2 structural proteins, such that said capsid encapsidate said vector DNA;    such that said second infectious papillomavirus pseudoviral particle differs from said first infectious papillomavirus pseudoviral particle in that said second is a different serotype from said first.