Cell type specific gene transfers using retroviral vectors containing antibody-envelope fusion proteins and wild-type envelope fusion proteins

Abstract

A method of infecting target cells with retroviral vector particles having target cell specificity. The retroviral vector particles have a chimeric envelope protein consisting of an antigen binding site of an antibody or another peptide, fused to the envelope protein the retroviral vector. The antigen binding site or other peptide disrupts the natural viral receptor binding site. The method includes producing the retroviral vector and contacting the vector with the target cell such that the vector is internalized by the cell.

What is claimed is:  
     
         1 . A method of infecting a cell having a selected antigen comprising: 
 (a) producing a retroviral vector having an envelope protein wherein a viral receptor portion is at least partially defined by a single chain antibody and said viral receptor portion binds to said selected antigen: and    (b) contacting said retroviral vector with the cell such that said viral particle or a portion thereof is internalized into the cell;    wherein the cell having the selected antigen is infected by said retroviral vector.    
     
     
         2 . The method of  claim 1  wherein said retroviral vector is produced using a plasmid comprising a nucleotide sequence encoding a single molecule comprising a portion of an envelope protein and a portion of a single chain antibody.  
     
     
         3 . The method of  claim 2  wherein said nucleotide sequence encodes a portion of the envelope protein of Spleen Necrosis Virus.  
     
     
         4 . The method of  claim 2  wherein the plasmid is that identified in FIG. 2 d  as pTC5.  
     
     
         5 . A method of infecting a cell having a selected antigen comprising: 
 (a) producing a retroviral vector having a targeting peptide fused to the envelope protein of said retroviral vector to form a targeting envelop; and    (b) contacting said retroviral vector with the cell such that said viral particle or a portion thereof is internalized into the cell;    wherein the cell having the selected antigen is infected by said retroviral vector.    
     
     
         6 . The method according to  claim 5  wherein said targeting peptide replaces or disrupts the natural viral receptor binding site.  
     
     
         7 . The method of  claim 5  wherein said targeting peptide is the antigen binding site of an antibody.  
     
     
         8 . The method of  claim 5  wherein said targeting peptide is the receptor binding peptide of another virus.  
     
     
         9 . The method of  claim 5  wherein said targeting peptide is a peptide that specifically binds to a specific receptor of the target.  
     
     
         10 . The method of  claim 5  wherein said retroviral vector particle is spleen necrosis virus.  
     
     
         11 . The method according to  claim 5  wherein said the targeting peptide is a single chain antibody against the hapten dinotrophenol (anti-DNP-scFv).  
     
     
         12 . The method according to  claim 5  wherein the targeting peptide is an antigen binding site directed against a cell-surface protein of the target cell.  
     
     
         13 . The method according to  claim 5  wherein the retroviral vector comprises a targeting envelope and a wild-type envelope.  
     
     
         14 . A cell type specific method for introducing genes into vertebrate cells using retroviral vectors which comprises administering to the cells a retroviral vector particle having target cell specificity which comprises a retroviral vector having an antigen binding site of an antibody fused to the envelope protein of the retroviral vector, wherein the antigen binding site of the antibody replaces the natural viral receptor binding site.  
     
     
         15 . The method according to  claim 14  wherein the retroviral particle is spleen necrosis virus.  
     
     
         16 . The method according to  claim 14  wherein the antibody is a single chain antibody against the hapten dinitrophenol.  
     
     
         17 . The method according to  claim 14  wherein the targeting peptide is an antigen binding site directed against a cell-surface protein of the target cell.  
     
     
         18 . The method according to  claim 14  wherein the targeting peptide is the receptor binding peptide of another virus.  
     
     
         19 . The method according to  claim 14  wherein the retroviral vector comprises a targeting envelope and a wild-type envelope.  
     
     
         20 . The method according to  claim 19  wherein the wild type envelope is derived from spleen necrosis virus.