Broadly reactive mosaic peptide for influenza vaccine

Abstract

The invention provides for mosaic influenza virus HA and NA sequences and uses thereof.

What is claimed is: 
     
         1 . A recombinant nucleic acid molecule having a) a nucleotide sequence encoding an immunogenic influenza virus HA polypeptide having one of SEQ ID NOs:1-11, a sequence with at least 99% amino acid sequence identity thereto, or a portion thereof, which provides cross-clade reactivity; b) a nucleotide sequence encoding an immunogenic H5 HA polypeptide having SEQ ID NO:1, or a sequence with at least 95% amino acid sequence identity thereto or an immunogenic portion thereof, wherein the HA has Ile at position 87, Thr at position 172, Val at position 226 or Thr at position 279, or a combination thereof; c) a nucleotide sequence encoding an immunogenic H1 HA polypeptide having SEQ ID NO:2, 3, 4, or 5, a sequence with at least 95% amino acid sequence identity thereto or an immunogenic portion thereof, wherein the HA i) has Arg at position 206, Leu at position 432, or Val at position 434, or a combination hereof; ii) has Ile at position 125 and Val at position 564; or iii) has Lys at position 62, Ile at position 64, Gln at position 68, Asn at position 71, Ser at position 73, Val at position 74, Leu at position 86, Ile at position 88, Ser at position 89, Lys at position 90, Glu at position 91, Lys at position 99, Pro at position 100, Asn at position 101, Pro at position 102, Glu at position 103, His at position 111, or Ala at position 113, or a combination thereof; d) a nucleotide sequence encoding an immunogenic H2 HA polypeptide having SEQ ID NO:6, a sequence with at least 95% amino acid sequence identity thereto or an immunogenic portion thereof, wherein the HA has Ala at position 24, Lys at position 45, Ser at position 87, Thr at position 258, Asn at position 260, or Leu at position 261, or a combination thereof; e) a nucleotide sequence encoding an immunogenic H7 HA polypeptide having SEQ ID NO:8, a sequence with at least 95% amino acid sequence identity thereto or an immunogenic portion thereof, wherein the HA has Ser at position 91, Ser at position 92, Arg at position 122, Gly at position 127, Glu at position 195, Val at position 197, or Ser at position 198, or a combination thereof; f) a nucleotide sequence encoding an immunogenic H9 HA polypeptide having SEQ ID NO:9, a sequence with at least 95% amino acid sequence identity thereto or an immunogenic portion thereof, wherein the HA has Gln at position 180, Glu at position 215, or Tyr at position 240, or a combination thereof; g) a nucleotide sequence encoding an influenza B HA polypeptide having SEQ ID NO:11, a sequence with at least 95% amino acid sequence identity thereto or an immunogenic portion thereof, wherein the HA has Met at position 86, Val at position 88, Thr at position 90, Thr at position 91, Lys at position 95, Ala at position 96, or Val at position 161, or a combination thereof; h) a nucleotide sequence encoding a H3 HA polypeptide having SEQ ID NO:7; i) a nucleotide sequence encoding a H10 HA polypeptide having having SEQ ID NO:10; j) a nucleotide sequence encoding an immunogenic influenza virus NA polypeptide having one of SEQ ID NOs:12-14; k) a sequence with at least 95% amino acid sequence identity to SEQ ID NO:12 having Ala at position 35, Ser at position 42, Asn at position 44, His at position 45, Thr at position 46, Gly at position 47, Ile at position 48, Arg at position 52, Ser at position 59, His at position 64, Asn at position 70, Val at position 74, Val at position 75, Ala at position 76, Gly at position 77, Asp at position 79, Lys at position 80, Thr at position 81, Ile at position 99, or Ser at position 105, or a combination thereof; I) a sequence with at least 99% amino acid sequence identity to SEQ ID NO:13 having Lys at position 199, Asn at position 221, or Gln at position 433, or a combination thereof; or m) a sequence with at least 99% amino acid sequence identity to SEQ ID NO:14 having Ile at position 353; or an immunogenic portion thereof. 
     
     
         2 . The recombinant nucleic acid molecule of  claim 1  wherein the nucleotide sequence is linked to a promoter operable in avian or mammalian cells. 
     
     
         3 . A vaccine comprising a recombinant virus, the genome of which comprises at least one expression cassette having a promoter operably linked to a heterologous open reading frame comprising a nucleotide sequence for an influenza virus polypeptide having one of SEQ ID NOs: 1-14, a polypeptide with at least 95% amino acid sequence thereto or an immunogenic portion thereof, or a combination thereof, which provides cross-clade reactivity. 
     
     
         4 . The vaccine of  claim 3  further comprising an adjuvant. 
     
     
         5 . The vaccine of  claim 3  further comprising a different virus. 
     
     
         6 . The vaccine of  claim 3  further comprising a pharmaceutically acceptable carrier. 
     
     
         7 . The vaccine of  claim 3  wherein the carrier is suitable for intranasal or intramuscular administration. 
     
     
         8 . The vaccine of  claim 3  which is in freeze-dried form. 
     
     
         9 . The vaccine of  claim 3  which is adapted for mucosal, intramuscular or intradermal delivery. 
     
     
         10 . A method to prevent, inhibit or treat influenza virus infection comprising administering to an animal or an egg thereof, a composition comprising an amount of at least one recombinant virus, the genome of which comprises at least one expression cassette having a promoter operably linked to the recombinant nucleic acid molecule of  claim 1 , effective to induce an adaptive immune response to influenza virus. 
     
     
         11 . The method of  claim 10  wherein the animal is an avian or a mammal. 
     
     
         12 . The method of  claim 10  wherein the composition is intradermally administered. 
     
     
         13 . The method of  claim 10  wherein the composition is intramuscularly administered. 
     
     
         14 . The method of  claim 10  wherein the composition is mucosally administered. 
     
     
         15 . The method of  claim 10  wherein the effective amount is administered in more than one dose. 
     
     
         16 . The method of  claim 10  wherein the composition further comprises an adjuvant. 
     
     
         17 . The method of  claim 10  wherein the composition is parenterally administered. 
     
     
         18 . The method of  claim 10  wherein the composition is administered intranasally. 
     
     
         19 . The method of  claim 10  where the composition is administered orally. 
     
     
         20 . The method of  claim 10  wherein the amount prevents or inhibits influenza virus infection across two or more clades.