Modifications of HIV Env, Gag, and Pol enhance immunogenicity for genetic immunization

Abstract

Modified HIV Env, Gag, Pol, or Nef DNA with improved ability to elicit antibody and CTL responses to HIV antigens have been identified as prototype immunogens for the treatment and prevention of HIV infections.

1 . A nucleic acid encoding a modified HIV Env glycoprotein wherein said modified HIV Env glycoprotein has a termination codon at position 704. 
     
     
         2 . The nucleic acid of  claim 1 , wherein said Env is from an R5 tropic HIV strain. 
     
     
         3 . The nucleic acid of  claim 1 , wherein said Env is from an X4 tropic HIV strain. 
     
     
         4 . The nucleic acid of  claim 1 , wherein the codons in said nucleic acid have been optimized for expression in humans. 
     
     
         5 . The nucleic acid of  claim 1 , wherein at least one variable region has been deleted. 
     
     
         6 . The nucleic acid of  claim 5 , wherein said at least one variable region is the V1 loop. 
     
     
         7 . The nucleic acid of  claim 6 , wherein the V2 loop has been deleted. 
     
     
         8 . The nucleic acid of  claim 5 , wherein said at least one variable region is selected from the group consisting of the V1 loop, the V2 loop, the V3 loop and the V4 loop. 
     
     
         9 . The nucleic acid of  claim 1 , wherein said modified HIV Env glycoprotein is from a lade selected from the group consisting of Clade A, Clade B and Clade C. 
     
     
         10 . The nucleic acid of  claim 9 , wherein said modified HIV Env glycoprotein is from Clade C (South African). 
     
     
         11 . The nucleic acid of  claim 1 , wherein said nucleic acid encodes a modified HIV Env glycoprotein in which the cleavage site, fusion domain, and at least part of the interspace between the two heptad-repeats are deleted. 
     
     
         12 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises the insert in SEQ ID NO: 92 which encodes a modified HIV Env glycoprotein. 
     
     
         13 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises a nucleotide sequence at least 95% identical to the nucleotide sequence of the insert in SEQ ID NO: 92 which encodes a modified HIV Env glycoprotein. 
     
     
         14 . The nucleic acid of  claim 1 , wherein said nucleic acid encodes a polypeptide at least 95% identical to the polypeptide encoded by the insert in SEQ ID NO: 92 which encodes a modified HIV Env glycoprotein. 
     
     
         15 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises the insert in SEQ ID NO: 14 which encodes a modified HIV Env glycoprotein. 
     
     
         16 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises a nucleotide sequence at least 95% identical to the nucleotide sequence of the insert in SEQ ID NO: 14 which encodes a modified HIV Env glycoprotein. 
     
     
         17 . The nucleic acid of  claim 1 , wherein said nucleic acid encodes a polypeptide at least 95% identical to the polypeptide encoded by the insert in SEQ ID NO: 14 which encodes a modified HIV Env glycoprotein. 
     
     
         18 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises the insert in SEQ ID NO: 96 which encodes a modified HIV Env glycoprotein. 
     
     
         19 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises a nucleotide sequence at least 95% identical to the nucleotide sequence of the insert in SEQ ID NO: 96 which encodes a modified HIV Env glycoprotein. 
     
     
         20 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises a nucleotide sequence at least 95% identical to the nucleotide sequence of the insert in SEQ ID NO: 96 which encodes a modified HIV Env glycoprotein. 
     
     
         21 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises the insert which encodes a modified HIV Env glycoprotein in a sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 954, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 86, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 142, SEQ ID NO: SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, and SEQ ID NO: 156 or a sequence at least 95% identical to the sequence of said insert. 
     
     
         22 . The nucleic acid of  claim 1 , wherein said nucleic acid comprises a nucleotide sequence encoding a polypeptide at least 95% identical to the polypeptide encoded by the insert which encodes a modified HIV Env glycoprotein in a sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 954, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 86, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 142, SEQ ID NO: SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, and SEQ ID NO: 156. 
     
     
         23 . The nucleic acid of  claim 1  further comprising a backbone, wherein said backbone is a plasmid vector. 
     
     
         24 . The nucleic acid of  claim 1  further comprising a backbone, wherein said backbone is an adenoviral vector. 
     
     
         25 . A nucleic acid which is at least 95% identical to the nucleic acid of  claim 1 , wherein said nucleic acid encodes a modified HIV Env glycoprotein with a termination codon at position 704. 
     
     
         26 . A modified HIV Env glycoprotein encoded by the nucleic acid molecule of  claim 1 . 
     
     
         27 . A vaccine composition comprising the nucleic acid molecule of  claim 1  and a pharmaceutically acceptable solution in a prophylactically effective dose. 
     
     
         28 . The composition of  claim 27  further comprising an adjuvant. 
     
     
         29 . A method of generating an antibody or CTL response against native HIV Env comprising the step of administering the nucleic acid molecule of  claim 1  to a host to generate said antibody or CTL response against native HIV Env. 
     
     
         30 . The method of  claim 29 , wherein said step is a primary immunization. 
     
     
         31 . The method of  claim 29 , wherein said step is a boost. 
     
     
         32 . The method of  claim 29 , wherein said step is a primary immunization with a plasmid vector. 
     
     
         33 . The method of  claim 29  wherein said step is a primary immunization with a plasmid vector expressing gp145delCFI and further comprising a step which is a boost with an adenoviral vector expressing gp140delCFI.