Adenovirus carrying gag gene HIV vaccine

Abstract

An adenoviral vector is described which carries a codon-optimized gag gene, along with a heterologous promoter and transcription terminator. This viral vaccine can effectively prevent HIV infection when administered to humans either alone or as part of a prime and boost regime also with a vaccine plasmid.

What is claimed is:  
     
         1 . A vaccine composition comprising a replication defective adenoviral vector comprising at least one gene encoding a HIV gag protein which is codon optimized for expression in a human, and the gene is operably linked to a heterologous promoter and transcription terminator.  
     
     
         2 . An adenoviral vaccine vector comprising: 
 a) a replication defective adenoviral vector, wherein the adenoviral vector does not have a functional E1 gene, and further comprising:    b) a gene expression cassette comprising: 
 i) a nucleic acid encoding a gag protein which is codon optimized for expression in a human host;  
 ii) a heterologous promoter operatively linked to the nucleic acid encoding the gag protein; and  
 iii) a transcription terminator.  
   
     
     
         3 . A vector according to  claim 2 , wherein the E1 gene has been deleted from the adenoviral vector.  
     
     
         4 . A vector according to  claim 3 , wherein the gene expression cassette has replaced the deleted E1 gene.  
     
     
         5 . A vector according to  claim 3 , wherein the adenovirus vector does not have a functional E3 gene.  
     
     
         6 . A vector according to  claim 5  wherein the E3 gene has been deleted from the replication defective adenoviral vector.  
     
     
         7 . A vector according to  claim 6  comprising adenoviral 5 sequences deleted of E1 region base pairs (bp) 342-3523 and deleted of E3 region bp 28,133-30,818.  
     
     
         8 . A vector according to  claim 6  comprising adenoviral 2 deleted of E1 region bp 559-3503 and E3 region bp 28,812-29,773.  
     
     
         9 . A vector according to  claim 8  comprising the sequence given in FIG. 6.  
     
     
         10 . A vector according to  claim 8  wherein the sequence is tPA-gag.  
     
     
         11 . A vector according to  claim 2  further comprising a physiologically acceptable carrier.  
     
     
         12 . An adenoviral vaccine composition for producing an immune response against human immunodeficiency virus (HIV) in a human comprising: 
 a) adenovirus serotype 5 sequences bp 1 to bp 341 and bp 3534 to 5798; and    b) a gene expression cassette, located 3′to adenovirus sequence bp 341, comprising: 
 i) a nucleic acid encoding gag which is codon-optimized and optionally has the tPA leader sequence at its 5′end;  
 ii) a human CMV promoter plus intron A operatively linked to the nucleic acid encoding gag; and  
 iii) a bovine growth hormone transcription terminator.  
   
     
     
         13 . A plasmid vector comprising: 
 a) an adenoviral portion comprising an adenoviral vector according to  claim 2;  and    b) a plasmid portion.    
     
     
         14 . A cell comprising an adenoviral vector of  claim 2 .  
     
     
         15 . A method of producing the vector of  claim 2  comprising introducing the adenoviral vector of  claim 2  into a host cell which expresses adenoviral E1 protein, and harvesting the resultant adenoviral vectors.  
     
     
         16 . A method according to  claim 15  wherein the cell is a 293 cell or PER.C6 cell.  
     
     
         17 . A method of generating a cellular immune response against an HIV protein in an individual comprising administering to the individual at least one adenovirus vaccine vector and a vaccine plasmid, 
 wherein said adenovirus vaccine vector comprises    a) a replication defective adenoviral vector, wherein the adenoviral vector does not have a functional E1 gene, and    b) a gene expression cassette comprising: i) a nucleic acid encoding gag protein optimized for expression in a human host; ii) a heterologous promoter operatively linked to the nucleic acid encoding the gag protein; and iii) a transcription terminator wherein said vaccine plasmid comprises: irus portion comprises    a) a gene expression cassette comprising: a nucleic acid encoding a gag protein, wherein the nucleic acid is codon optimized for expression in a human host;    b) a promoter; and    c) a transcription terminator    wherein the vaccine plasmid does not contain any adenoviral genes.    
     
     
         18 . A method according to  claim 17  comprising administering a vaccine plasmid to the individual, and after a predetermined minimum amount of time has passed, administering an adenovirus vaccine vector to the individual.  
     
     
         19 . A method according to  claim 17  comprising administering an adenovirus vaccine vector to the individual, and after a predetermined minimum amount of time has passed, administering a vaccine plasmid to the individual.  
     
     
         20 . A method according to  claim 14  comprising administering an adenovirus vaccine vector to the individual, and after a predetermined minimum amount of time has passed, re-administering an adenovirus vector to the individual.