Conditionally replicating viral vectors and their use

Abstract

The present invention provides a conditionally replicating viral vector, methods of making, modifying, propagating and selectively packaging, and using such a vector, isolated molecules of specified nucleotide and amino acid sequences relevant to such vectors, a pharmaceutical composition and a host cell comprising such a vector, the use of such a host cell to screen drugs. The methods include the prophylactic and therapeutic treatment of viral infection, in particular HIV infection, and, thus, are also directed to viral vaccines and the treatment of cancer, in particular cancer of viral etiology. Other methods include the use of such conditionally replicating viral vectors in gene therapy and other applications.

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         29 . A method of stimulating an immune response in a human subject, wherein said method comprises 
 contacting a cell of said subject with a conditionally replicating human immunodeficiency viral (crHIV) vector comprising a first nucleotide sequence, wherein said contacting occurs ex vivo, and    stimulating an immune response in said subject upon conditional replication of said crHIV after infection of said cell by wild-type HIV virus or after introduction of a helper vector,    wherein said first sequence adversely affects said HIV; and    wherein said crHIV expresses one or more HIV proteins necessary for replication of said crHIV vector or packaging said crHIV into a virion; and    wherein said crHIV viral vector replicates in a host cell only upon complementation with a wild-type virus or a helper virus, or a helper vector, and wherein said complementation renders the host cell permissive for replication of said crHIV vector; and    wherein said crHIV vector is selectively replicated over said wild-type virus, helper virus, or helper vector.    
     
     
         30 . The method of  claim 29  wherein said conditional replication of crHIV results in a persistent expression of said one or more HIV proteins necessary for replication of said crHIV vector.  
     
     
         31 . The method of  claim 29  wherein said conditional replication of crHIV results in a persistent expression of said one or more HIV proteins necessary for packaging of said crHIV vector into a virion.  
     
     
         32 . The method of  claim 29  wherein said first nucleotide sequence comprises or encodes, in which case it also expresses, a genetic antiviral agent.  
     
     
         33 . The method of  claim 32  wherein said genetic antiviral agent is an antisense molecule, a ribozyme, a nucleic acid decoy, a transdominant mutant protein, a single chain antibody, a cytokine, an antigen, a receptor, or a suicide gene.  
     
     
         34 . The method of  claim 33  wherein said genetic antiviral agent is a cytokine, an antigen, or a suicide gene.  
     
     
         35 . The method of  claim 29  wherein said first nucleotide sequence encodes a mutated HIV protein.  
     
     
         36 . The method of  claim 29  wherein said first nucleotide sequence is operably linked to the LTR of said crHIV vector.  
     
     
         37 . The method of  claim 29  wherein said cell of a subject is infected with HIV before said contacting with a crHIV vector.  
     
     
         38 . The method of  claim 29  wherein said cell of a subject is not infected with HIV before said contacting with a crHIV vector.  
     
     
         39 . The method of  claim 28  further comprising introducing a helper vector into said cell.  
     
     
         40 . The method of  claim 29  wherein said helper vector is replicated in said cell and/or packaged into a virion via complementation by said crHIV vector.  
     
     
         41 . The method of  claim 40  wherein said helper vector comprises a nucleotide sequence that reduces recombination with said crHIV vector.  
     
     
         42 . The method of  claim 41  wherein said nucleotide sequence that reduces recombination with said crHIV vector is a ribozyme.  
     
     
         43 . The method of  claim 29  wherein said cell of a subject is an antigen presenting cell or a T cell.  
     
     
         44 . The method of  claim 43  wherein said cell of a subject is an antigen presenting cell.  
     
     
         45 . The method of  claim 43  wherein said cell of a subject is a T cell.  
     
     
         46 . The method of  claim 29  wherein said crHIV vector is packaged in an infectious viral particle or formulated in a liposome or with an adjuvant.  
     
     
         47 . The method of  claim 29  wherein said crHIV vector is a chimeric vector comprising sequences derived from different viruses.  
     
     
         48 . A pair of vectors, said pair comprising a conditionally replicating human immunodeficiency viral (crHIV) vector comprising a first nucleotide sequence and 
 a helper vector which complements replication of said crHIV vector,    wherein said first sequence adversely affects wild-type HIV virus and said helper vector; and    wherein said crHIV viral vector replicates in a host cell only upon complementation with a wild-type virus or a helper virus, or a helper vector, and wherein said complementation renders the host cell permissive for replication of said crHIV vector; and    wherein said crHIV vector is selectively replicated over said wild-type virus, helper virus, or helper vector.    
     
     
         49 . The pair of  claim 48  wherein said helper vector comprises a nucleotide sequence that reduces recombination with said crHIV vector.  
     
     
         50 . The pair of  claim 49  wherein said nucleotide sequence that reduces recombination with said crHIV vector is a ribozyme.  
     
     
         51 . The pair of  claim 48  wherein said first nucleotide sequence comprises or encodes, in which case it also expresses, a genetic antiviral agent.  
     
     
         52 . The pair of  claim 51  wherein said genetic antiviral agent is an antisense molecule, a ribozyme, a nucleic acid decoy, a transdominant mutant protein, a single chain antibody, a cytokine, an antigen, a receptor, or a suicide gene.  
     
     
         53 . The pair of  claim 48  wherein said first nucleotide sequence encodes a mutated HIV protein.  
     
     
         54 . The pair of  claim 48  wherein one or both of said vectors are packaged in an infectious viral partial or formulated in a liposome or with an adjuvant.  
     
     
         55 . The pair of  claim 48  wherein said crHIV vector is a chimeric vector comprising sequences derived from different viruses.