Polynucleotides encoding antigenic hiv type b polypeptides, polypeptides, and uses thereof

Abstract

The present invention relates to polynucleotides encoding immunogenic HIV polypeptides. Uses of the polynucleotides in applications including immunization, generation of packaging cell lines, and production of HIV polypeptides are also described. Polynucleotides encoding antigenic HIV polypeptides are described, as are uses of these polynucleotides and polypeptide products therefrom, including formulations of immunogenic compositions and uses thereof.

1 . A nucleic acid comprising an expression cassette, wherein the expression cassette comprises a polynucleotide sequence encoding a polypeptide including a polypeptide selected from the group consisting of: 
 (a) an HIV Gag polypeptide, wherein the polynucleotide sequence encoding said Gag polypeptide comprises a sequence having at least 90% sequence identity to the full length of a sequence selected from the group consisting of SEQ ID NO:9; SEQ ID NO:10; SEQ ID NO:11; SEQ ID NO:12; and SEQ ID NO:16;    (b) an HIV Gag polypeptide, wherein the polynucleotide sequence encoding said Gag polypeptide comprises a sequence having at least 98% sequence identity to the full length of a sequence selected from the group consisting of SEQ ID NO:13; SEQ ID NO:14; SEQ ID NO:15; and SEQ ID NO:55;    (c) an HIV Gag polypeptide, wherein the polynucleotide sequence encoding said Gag polypeptide comprises a sequence having at least 95% sequence identity to the full length of SEQ ID NO:17;    (d) an HIV Nef polypeptide, wherein the polynucleotide sequence encoding said Nef polypeptide comprises a sequence having at least 90% sequence identity to the full length of a sequence selected from the group consisting of SEQ ID NO:25; SEQ ID NO:26; SEQ ID NO:27; SEQ ID NO:28; SEQ ID NO:33 and SEQ ID NO:34;    (e) an HIV Prot polypeptide, wherein the polynucleotide sequence encoding said Prot polypeptide comprises a sequence having at least 98% sequence identity to the full length of SEQ ID NO:39;    (f) an HIV Tat polypeptide, wherein the polynucleotide sequence encoding said Tat polypeptide comprises a sequence having at least 90% sequence identity to the full length of a sequence selected from the group consisting of SEQ ID NO:42; SEQ ID NO:43; SEQ ID NO:51; SEQ ID NO:53; SEQ ID NO:54; SEQ ID NO:56; and SEQ ID NO:57;    (g) an HIV Rev polypeptide, wherein the polynucleotide sequence encoding said Rev polypeptide comprises a sequence having at least 90% sequence identity to the full length of SEQ ID NO:44 or SEQ ID NO:45;    (h) an HIV Vif polypeptide, wherein the polynucleotide sequence encoding said Vif polypeptide comprises a sequence having at least 90% sequence identity to at least 30 contiguous base pairs of SEQ ID NO:58;    (i) an HIV Vpr polypeptide, wherein the polynucleotide sequence encoding said Vpr polypeptide comprises a sequence having at least 90% sequence identity to at least 20 contiguous base pairs of SEQ ID NO:59;    (j) an HIV Vpu polypeptide, wherein the polynucleotide sequence encoding said Vpu polypeptide comprises a sequence having at least 90% sequence identity to at least 20 contiguous base pairs of SEQ ID NO:60; and    (k) an HIV Env polypeptide, wherein the polynucleotide sequence encoding said Env polypeptide comprises a sequence having at least 90% sequence identity to the full length of a sequence selected from the group consisting of SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, and SEQ ID NO:64.    
     
     
         2 . The nucleic acid of  claim 1  which is a recombinant expression system for use in a selected host cell, wherein the recombinant expression system comprises an expression cassette of  claim 1 , wherein said polynucleotide sequence is operably linked to control elements compatible with expression of the polynucleotide in the selected host cell.  
     
     
         3 . The recombinant expression system of  claim 2  wherein said control elements are selected from the group consisting of a transcription promoter, a transcription enhancer element, a transcription termination signal, polyadenylation sequences, sequences for optimization of initiation of translation, and translation termination sequences.  
     
     
         4 . The recombinant expression system of  claim 2  wherein said transcription promoter is selected from the group consisting of CMV, CMV+intron A, SV40, RSV, HIV-Ltr, MMLV-ltr, and metallothionein.  
     
     
         5 . An isolated cell comprising an expression cassette of  claim 1 , wherein said polynucleotide sequence is operably linked to control elements compatible with expression in the selected cell.  
     
     
         6 . The isolated cell of  claim 5  which is selected from the group consisting of a mammalian cell, an insect cell, a bacterial cell, a yeast cell, a plant cell, an antigen presenting cell, a primary cell, an immortalized cell, and a tumor-derived cell.  
     
     
         7 . A method for producing a polypeptide including HIV polypeptide sequences, comprising incubating isolated cells of  claim 5  under conditions for producing said polypeptide.  
     
     
         8 . The nucleic acid of  claim 1  which is a vector for use in generating an immune response in a mammalian subject, wherein the vector comprises an expression cassette of  claim 1 , and wherein said polynucleotide sequence is operably linked to control elements compatible with expression of the polynucleotide in the subject.  
     
     
         9 . A method of DNA immunization of a subject, comprising, introducing a vector of  claim 8  into said subject under conditions that are compatible with expression of said expression cassette in said subject.  
     
     
         10 . The method of  claim 9 , wherein said vector is: 
 (a) a nonviral vector;    (b) encapsulated in a liposome preparation; or    (c) a viral vector.    
     
     
         11 . The method of  claim 9  wherein said vector is delivered using a particulate carrier.  
     
     
         12 . The method of  claim 11  wherein said vector is coated on a gold or tungsten particle and said coated particle is delivered to said subject using a gene gun.  
     
     
         13 . The method of  claim 9  wherein said vector is a viral vector and the viral vector is selected from the group consisting of a retroviral vector, an alphaviral vector, and a lentiviral vector.  
     
     
         14 . The method of  claim 9  wherein said subject is a mammal.  
     
     
         15 . The method of  claim 14  wherein said mammal is a human.  
     
     
         16 . The method of  claim 9  wherein cells of the subject are transfected with the vector of  claim 10  under conditions that permit the expression of said polynucleotide and production of said polypeptide, thereby eliciting an immunological response to said polypeptide.  
     
     
         17 . The method of  claim 16  wherein said transfecting is done ex vivo and said transfected cells are reintroduced into said subject.  
     
     
         18 . The method of  claim 16  wherein said transfecting is done in vivo in said subject.  
     
     
         19 . The method of  claim 16  where said immune response is a humoral immune response.  
     
     
         20 . The method of  claim 16  where said immune response is a cellular immune response.  
     
     
         21 . The method of  claim 16  wherein the vector is administered intramuscularly, intramucosally, intranasally, subcutaneously, intradermally, transdermally, intravaginally, intrarectally, orally or intravenously.