HIV envelopolype peptides

Abstract

A method for the rational design and preparation of vaccines based on HIV envelope polypeptides is described. In one embodiment, the method for making an HIV gp120 subunit vaccine for a geographic region comprises determining neutralizing epitopes in the V2 and/or C4 domains of gp120 of HIV isolates from the geographic region and selecting an HIV strain having gp120 a neutralizing epitope in the V2 or C4 domain which is common among isolates in the geographic region. In a preferred embodiment of the method, neutralizing epitopes for the V2, V3, and C4 domains of gp120 are determined. At least two HIV isolates having different neutralizing epitopes in the V2, V3, or C4 domain are selected and used to make the vaccine. The invention also provides a multivalent HIV gp120 subunit vaccine. A DNA sequence encoding gp120 from preferred vaccine strains of HIV, GNE 8 and GNE 16 , expression constructs comprising the GNE 8 -gp120 and GNE 16 -gp120 encoding DNA under the transcriptional and translational control of a heterologous promoter, and isolated GNE 8 -gp120 and GNE 16 -gp120 are also described.

What is claimed is:  
     
         1 . A method for making On HIV gp120 subunit vaccine for a geographic region comprising the steps of: 
 a determining a neutralizing epitope in the V2 or C4 domain of gp120 of HIV isolates from the geographic region;    b. selecting an HIV strain having gp120 which has a neutralizing epitope in the V2 or C4 domain which is common among isolates in the geographic region; and    c. making an HIV gp120 subunit vaccine from the selected isolate.    
     
     
         2 . The method of  claim 1  wherein the neutralizing epitope is determined by determining the amino acid sequence for at least a portion of the V2 or C4 domain.  
     
     
         3 . The method of  claim 2  wherein the amino acid sequence is determined by sequencing DNA encoding at least a portion of the V2 or C4 domain.  
     
     
         4 . The method of  claim 3  wherein a plurality of isolates having different amino acid sequences for the V2 and C4 domains are selected.  
     
     
         5 . The method of  claim 4  wherein a plurality of isolates having different amino acid sequences for the V3 domain is selected.  
     
     
         6 . A method for making an HIV gp120 subunit vaccine for a geographic region comprising the steps of: 
 a. determining neutralizing epitopes for the V2, V3, and C4 domains of gp120 from HIV isolates from the geographic region;    b. selecting at least two HIV isolates having different neutralizing epitopes in the V2, V3, or C4 domain; and    c. making an HIV gp120 subunit vaccine from the selected isolates.    
     
     
         7 . The method of  claim 6  wherein each of the selected isolates have one of the most common neutralizing epitopes.  
     
     
         8 . A method for making an HIV gp120 subunit vaccine for a geographic region comprising the steps of: 
 a. determining the neutralizing epitopes for HIV isolates and the percentage of HIV infections attributable to each strain present in the region;    b. selecting at least two HIV strains which have the most common neutralizing epitopes in the V2, V3, and C4 domains in the geographic region; and    c. making an HIV gp120 subunit vaccine from the selected isolates.    
     
     
         9 . The method of  claim 8  wherein the isolates are primary patient isolates.  
     
     
         10 . The method of  claim 8  wherein the geographic region is North America and the amino acid sequence of gp120 from the HIV isolates MN and GNE 8  are selected.  
     
     
         11 . The method of  claim 10  wherein the GNE 16  isolate is also selected.  
     
     
         12 . A multivalent HIV gp120 subunit vaccine.  
     
     
         13 . The vaccine of  claim 12  wherein gp120 present in the vaccine is from at least two HIV isolates which have a different neutralizing epitope in the V2 or C4 domain of gp120.  
     
     
         14 . The vaccine of  claim 12  wherein gp120 present in the vaccine is from at least two HIV isolates which have a different neutralizing epitope in the V3 domain of gp120.  
     
     
         15 . The vaccine of  claim 12  wherein each isolate has a different common neutralizing epitope for the V2 or C4 domains of gp120.  
     
     
         16 . The vaccine of  claim 1  wherein gp120 present in the vaccine is from the MN and GNE 8  strains of HIV.  
     
     
         17 . The vaccine of  claim 1  wherein gp120 from the GNE 16  strain of HIV is also present in the vaccine.  
     
     
         18 . A DNA sequence of less than 5 kilobases encoding gp120 from GNE$ and having the nucleotide sequence illustrated in Table 1.  
     
     
         19 . A DNA sequence of less than 5 kilobases encoding gp120 from GNE 16  and having the nucleotide sequence illustrated in Table 2.  
     
     
         20 . An expression construct comprising DNA encoding gp120 selected from the group consisting of GNE 8 -gp120 and GNE 16 -gp120 under the transcriptional and translational control of a heterologous promoter.  
     
     
         21 . The expression construct of  claim 20  wherein the promoter is a eukaryotic promoter.  
     
     
         22 . The expression construct of  claim 21  wherein the DNA encoding gp120 is joined to a heterologous signal sequence.  
     
     
         23 . An isolated GNE,-gp120 polypeptide having the amino acid sequence illustrated in Table 1.  
     
     
         24 . An isolated GNE 16 -gp120 polypeptide having the amino acid sequence illustrated in Table 2.  
     
     
         25 . An improved serotyping method for HIV strains comprising determining the serotypes of the V2, V3, and C4 domains of gp120.  
     
     
         26 . A truncated gp120 sequence which sequence is free from the C5 domain.  
     
     
         27 . The truncated gp120 sequence of  claim 26  wherein the sequence is additionally free from the carboxy terminus through amino acid residue 453 of the gp120 V5 domain.  
     
     
         28 . The truncated gp120 sequence of  claim 27  wherein the sequence is additionally free from the gp120 V5 domain.  
     
     
         29 . The truncated gp120 sequence of  claim 26  wherein the sequence is additionally free from the gp120 signal sequence.  
     
     
         30 . The truncated gp120 sequence of  claim 29  wherein the sequence is additionally free from the carboxy terminus through amino acid residue 111 of the gp120 C1 domain.  
     
     
         31 . The truncated gp120 Sequence of  claim 29  wherein the sequence is additionally free from the carboxy terminus through amino acid residue 117 of the gp120 C1 domain.  
     
     
         32 . The truncated gp120 sequence of  claim 26  wherein the sequence is free from the amino terminus of gp120 through residue 111 of the C1 domain and residue 453 through the carboxy terminus of gp120.  
     
     
         33 . The truncated gp120 sequence of  claim 26  wherein the sequence is produced by recombinant engineering.