Influenza virus vaccine composition and methods of use

Abstract

The present invention is directed to enhancing the immune response of a human in need of protection against IV infection by administering in vivo, into a tissue of the human, at least one polynucleotide comprising one or more regions of nucleic acid encoding an IV protein or a fragment, a variant, or a derivative thereof. The present invention is further directed to enhancing the immune response of a human in need of protection against IV infection by administering, in vivo, into a tissue of the human, at least one IV protein or a fragment, a variant, or derivative thereof. The IV protein can be, for example, in purified form or can be an inactivated IV, such as those present in inactivated IV vaccines. The polynucleotide is incorporated into the cells of the human in vivo, and an immunologically effective amount of an immunogenic epitope of an IV, or a fragment, variant, or derivative thereof is produced in vivo. The IV protein (in purified form or in the form of an inactivated IV vaccine) is also administered in an immunologically effective amount.

1 - 392 . (canceled)  
     
     
         393 . An isolated polynucleotide comprising a nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:78, wherein the codons of said nucleic acid fragment are optimized for expression in humans.  
     
     
         394 . The polynucleotide of  claim 393 , wherein the nucleotide sequence of said nucleic acid fragment is SEQ ID NO:66.  
     
     
         395 . A vector comprising the polynucleotide of  claim 393 , wherein said vector, upon uptake by a suitable host cell, expresses said amino acid sequence.  
     
     
         396 . The polynucleotide of  claim 393 , further comprising a heterologous nucleic acid ligated to said nucleic acid fragment.  
     
     
         397 . A composition comprising the vector of  claim 395  and a carrier.  
     
     
         398 . The composition of  claim 397 , further comprising a component selected from the group consisting of an adjuvant and a transfection facilitating compound.  
     
     
         399 . The composition of  claim 398 , wherein said component is a cationic lipid.  
     
     
         400 . The composition of  claim 399 , wherein said adjuvant comprises(±)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(syn-9-tetradeceneyloxy)-1-propanaminium bromide (GAP-DMORIE) and a neutral lipid, wherein said neutral lipid is selected from the group consisting of: 
 (a) 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);    (b) 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (DPyPE); and    (c) 1,2-dimyristoyl-glycer-3-phosphoethanolamine (DMPE).    
     
     
         401 . The composition of  claim 399 , wherein said transfection facilitating compound comprises (±)-N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (DMRIE).  
     
     
         402 . The composition of  claim 401 , wherein said transfection facilitating compound further comprises a neutral lipid.  
     
     
         403 . The composition of  claim 402 , wherein said neutral lipid is DOPE.  
     
     
         404 . The composition of  claim 401  further comprising a 1:1 molar ratio of GAP-DMORIE and DPyPE.  
     
     
         405 . A method for treating or preventing influenza infection in a vertebrate comprising administering to a vertebrate in need thereof the composition of  claim 397 .  
     
     
         406 . A method for eliciting an immune response to influenza virus in a vertebrate comprising administering to a vertebrate in need thereof the composition of  claim 397 .  
     
     
         407 . An isolated polynucleotide comprising a nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:76, wherein the codons of said nucleic acid fragment are optimized for expression in humans.  
     
     
         408 . The polynucleotide of  claim 407 , wherein the nucleotide sequence of said nucleic acid fragment is SEQ ID NO:75.  
     
     
         409 . A vector comprising the polynucleotide of  claim 405 , wherein said vector, upon uptake by a suitable host cell, expresses said amino acid sequence.  
     
     
         410 . The polynucleotide of  claim 407 , further comprising a heterologous nucleic acid ligated to said nucleic acid fragment.  
     
     
         411 . A composition comprising the vector of  claim 409  and a carrier  
     
     
         412 . The composition of  claim 411 , further comprising a component selected from the group consisting of an adjuvant and a transfection facilitating compound.  
     
     
         413 . The composition of  claim 412 , wherein said component is a cationic lipid.  
     
     
         414 . The composition of  claim 413 , wherein said adjuvant comprises(±)-N-(3 -aminopropyl)-N,N-dimethyl-2,3-bis(syn-9-tetradeceneyloxy)-1-propanaminium bromide (GAP-DMORIE) and a neutral lipid, wherein said neutral lipid is selected from the group consisting of: 
 (a) 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);    (b) 1,2-diphytanoyl-sn-glycero-3 -phosphoethanolamine (DPyPE); and    (c) 1,2-dimyristoyl-glycer-3-phosphoethanolamine (DMPE).    
     
     
         415 . The composition of  claim 413 , wherein said transfection facilitating compound comprises (±)-N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (DMRIE).  
     
     
         416 . The composition of  claim 415 , wherein said transfection facilitating agent further comprises a neutral lipid.  
     
     
         417 . The composition of  claim 416 , wherein the neutral lipid is DOPE.  
     
     
         418 . The composition of  claim 415  further comprising a 1:1 molar ratio of GAP-DMORIE and DPyPE.  
     
     
         419 . An isolated polypeptide produced by the polynucleotide  claim 407 .  
     
     
         420 . A method for treating or preventing influenza infection in a vertebrate comprising administering to a vertebrate in need thereof the composition of  claim 411 .  
     
     
         421 . A method for eliciting an immune response to influenza virus in a vertebrate by administration of the composition of  claim 411 .  
     
     
         422 . An isolated polynucleotide comprising a first nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:78 and a second nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:76, wherein the codons of said first and second nucleic acid fragments are optimized for expression in humans.  
     
     
         423 . The polynucleotide of  claim 422 , wherein the nucleotide sequence of said first nucleic acid fragment is SEQ ID NO:66 and wherein the nucleotide sequence of said second nucleic acid fragment is SEQ ID NO:75.  
     
     
         424 . A vector comprising the polynucleotide of  claim 422 , wherein said vector, upon uptake by a suitable host cell, expresses the consensus amino acid sequences of SEQ ID NO:78 and SEQ ID NO:76.  
     
     
         425 . The vector of  claim 424 , wherein said consensus amino acid sequences of SEQ ID NO:78 and SEQ ID NO:76 are expressed as a fusion protein.  
     
     
         426 . The vector of  claim 422 , wherein said vector is DNA and wherein said vector comprises a first expression cassette and second expression cassette, said first expression cassette comprises a first nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:78 in operable association with a promoter and said second expression cassette comprises a second nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:76 in operable association with a promoter.  
     
     
         427 . The vector of  claim 426 , wherein said first expression cassette and said second expression cassette are associated with separate promoters.  
     
     
         428 . The vector of  claim 427 , wherein said separate promoters are non-identical.  
     
     
         429 . The vector of  claim 426 , wherein said first expression cassette and said second expression cassette are associated with a single promoter, and wherein said second expression cassette is in operable association with an internal ribosome entry site (IRES).  
     
     
         430 . The vector of  claim 426 , wherein said first expression cassette and said second expression cassette are associated with a single promoter, and wherein said first expression cassette is in operable association with an internal ribosome entry site (IRES).  
     
     
         431 . A composition comprising the vector of  claim 424  and a carrier.  
     
     
         432 . A composition comprising the vector of  claim 426  and a carrier.  
     
     
         433 . A composition comprising at least two non-identical vectors, wherein one of said vectors comprises a nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:78 and wherein another of said vectors comprises a nucleic acid fragment which encodes the consensus amino acid sequence of SEQ ID NO:76, wherein the codons of said nucleic acid fragments encoding SEQ ID NO:78 and SEQ ID NO:76 are optimized for expression in humans, and wherein said vectors, upon uptake by a suitable host cell, express said amino acid sequences.  
     
     
         434 . The composition of  claim 433 , further comprising a carrier.  
     
     
         435 . The composition of  claim 434 , further comprising a component selected from the group consisting of an adjuvant and a transfection facilitating compound.  
     
     
         436 . The composition of  claim 435 , wherein said component is a cationic lipid.  
     
     
         437 . The composition of  claim 436 , wherein said adjuvant comprises(±)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(syn-9-tetradeceneyloxy)-1-propanaminium bromide (GAP-DMORIE) and a neutral lipid, wherein said neutral lipid is selected from the group consisting of: 
 (a) 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);    (b) 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (DPyPE); and    (c) 1,2-dimyristoyl-glycer-3-phosphoethanolamine (DMPE).    
     
     
         438 . The composition of  claim 436 , wherein said transfection facilitating compound comprises (±)-N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (DMRIE).  
     
     
         439 . The composition of  claim 438 , wherein said transfection facilitating compound further comprises a neutral lipid.  
     
     
         440 . The composition of  claim 439 , wherein the neutral lipid is DOPE.  
     
     
         441 . The composition of  claim 438  further comprising a 1:1 molar ratio of GAP-DMORIE and DPyPE.  
     
     
         442 . A method for treating or preventing influenza infection in a vertebrate comprising administering to a vertebrate in need thereof the composition of  claim 434 .  
     
     
         443 . A method for eliciting an immune response to influenza virus in a vertebrate by administration of the composition of  claim 434.