Methods and compositions for impairing multiplication of HIV-1

Abstract

A composition which elicits antibodies to greater than 95%, and even greater than 99%, of the known variants of HIV-1 Tat protein contains at least one peptide or polypeptide of the formula of Epitope I (based on amino acids 2-10 of HIV-1 Tat consensus sequence) and optionally one or more of a peptide or polypeptide of Epitope II (based on amino acids 41 to 51 of that sequence), of Epitope III (based on amino acids 52-62 of that sequence), or of Epitope IV (based on amino acids 62 through 72 of that sequence with a C-terminal Pro). Vaccinal and pharmaceutical compositions can contain the antibodies induced by the peptide compositions for use in passive therapy. Diagnostic compositions and uses are described for assessing the immune status of vaccinated patients.

1 . An immunogenic composition comprising at least variant peptides, each peptide having the formula: Val-Asp-Pro-Y-Leu-Glu-Pro-Trp-Lys-His-Pro-Gly-Ser SEQ ID NO: 58, wherein each variant has a different amino acid at Y, said Y being selected from the group consisting of Arg, Lys, Ser and Asn, wherein said composition induces antibodies that bind to HIV-1 Tat proteins from different HIV-1 strains and subtypes.  
     
     
         2 . The composition according to  claim 1 , wherein said peptides are fused in frame in a single linear protein.  
     
     
         3 . The composition according to  claim 1 , wherein said peptides are each coupled to carrier protein.  
     
     
         4 . A composition comprising a nucleic acid sequence encoding at least two variants of the amino acid sequence R1-Val-Asp-Pro-Y-Leu-Glu-Pro-R2 SEQ ID NO: 36, wherein each variant has a different amino acid at Y, said Y being selected from the group consisting of Arg, Lys, Ser and Asn, wherein R1 is a sequence of up to 5 amino acids, wherein R2 is a sequence of up to 14 additional amino acids, or a fragment thereof, wherein said amino acid sequences or fragments induce antibodies that bind to HIV-1 Tat proteins from different HIV-1 strains and subtypes.  
     
     
         5 . The composition according to  claim 4 , wherein at least one said amino acid sequence is R1-Val-Asp-Pro-Arg-Leu-Glu-Pro-R2, R1-Val-Asp-Pro-Lys-Leu-Glu-Pro-R2, R1-Val-Asp-Pro-Ser-Leu-Glu-Pro-R2, or R1-Val-Asp-Pro-Asn-Leu-Glu-Pro-R2.  
     
     
         6 . The composition according to  claim 4 , wherein R1 is —X-Pro-, wherein X is selected from the group consisting of Glu or Asp.  
     
     
         7 . The composition according to  claim 4 , wherein R2 is selected from the group consisting of: -Trp, -Trp-Lys; -Trp-Lys-His; -Trp-Lys-His-Pro; -Trp-Lys-His-Pro-Gly; and -Trp-Lys-His-Pro-Gly-Ser-.  
     
     
         8 . The composition according to  claim 4 , wherein said nucleic acid sequence encodes said selected amino acid variant sequences in an open reading frame with, or fused to, at least one third nucleic acid sequence.  
     
     
         9 . The composition according to  claim 8 , wherein said third amino acid sequence is selected from the group consisting of a second copy of one of said at least two variant amino acid sequences or fragments, a third immunogenic amino acid sequence having a different sequence than said two variant amino acid sequences, and a carrier protein.  
     
     
         10 . The composition according to  claim 8 , wherein said third amino acid sequence is selected from at least one of the group consisting of: 
 (a) the amino acid sequence R3-Lys-X-Leu-Gly-Ile-Ser-Tyr-Gly-Arg-Lys-R4 of SEQ ID NO:37, wherein said amino acid X is Gly or Ala; wherein R3 is an optional sequence of between 1 to about 5 additional amino acids; and wherein R4 is an optional sequence of between about 1 to about 5 additional amino acids; or a fragment thereof;    (b) at least one amino acid sequence of SEQ ID NOS: 11-18;    (c) an amino acid sequence of R5-Arg-Arg-X-Z-A-Y-Ser- SEQ ID NO: 38, wherein X is selected from the group consisting of Ala, Pro, Ser, and Gln; wherein Y is selected from the group consisting of Asp, Asn, Gly and Ser; wherein Z is selected from the group consisting of Pro and His; wherein A is selected from the group consisting of Gln and Pro; and wherein R5 is an optional sequence of between 1 and 3 amino acids; and    (d) an amino acid sequence of R7-Ser-Gln-X-His-Gln-Y-Ser-Leu-Ser-Lys-Gln-Pro-R8 SEQ ID NO: 39, wherein X is selected from the group consisting of Asn and Thr; wherein Y is selected from the group consisting of Ala and Val; wherein R7 is an optional sequence of between 1 to 3 amino acids; and wherein R8 is a sequence of about 1 to 3 amino acids.    
     
     
         11 . The composition according to  claim 4 , wherein said nucleic acid sequence encodes sequentially at least three said variant amino acid sequences or fragments thereof, each variant having a different amino acid at position Y.  
     
     
         12 . The composition according to  claim 4 , wherein said nucleic acid sequence encodes sequentially at least four said variant amino acid sequences or fragments thereof, each variant having a different amino acid at position Y.  
     
     
         13 . The composition according to  claim 4 , wherein said nucleic acid sequence encodes multiple copies of the same amino acid sequence.  
     
     
         14 . A method of inducing an immune response in a mammal comprising administering to said mammal an effective amount of a composition of  claim 4 .  
     
     
         15 . A method of inducing an immune response in a mammal comprising administering to said mammal an effective amount of a composition of  claim 4 .  
     
     
         16 . An antibody composition comprising at least two different ISOLATED antibodies that bind all of the peptides of the composition of  claim 4 .  
     
     
         17 . An antibody composition comprising at least two different antibodies, said composition specifically binding to a least two variants of an epitope of an HIV-1 Tat protein, said epitope located within the amino acid sequence R1-Val-Asp-Pro-Y-Leu-Glu-Pro-R2 SEQ ID NO: 36, wherein Y is selected from the group consisting of Arg, Lys, Ser and Asn, and each variant has a different amino acid at Y, R1 is a sequence of up to 5 amino acids, R2 is a sequence of up to 14 additional amino acids, or a fragment thereof, and wherein said composition binds to HIV-1 Tag proteins from multiple different HIV-1 strains and subtypes.  
     
     
         18 . The composition according to  claim 17 , wherein each of said antibodies binds a different Y variant.  
     
     
         19 . The composition according to  claim 17 , comprising three different antibodies.  
     
     
         20 . The composition according to  claim 17 , comprising four different antibodies.