Recombinant Nucleic Acid Construct Encoding Influenza Peptides and Compositions

Abstract

The present specification discloses recombinant nucleic acid constructs encoding an immunogenic multiepitope polypeptide comprising two or more polypeptides, recombinant nucleic acid constructs encoding at least two epitopes from two or more internal proteins of influenza virus, compositions comprising such recombinant nucleic acid constructs and methods of eliciting a T cell immune response against an influenza virus in a vertebrate using such recombinant nucleic acid constructs and compositions.

1 . A recombinant nucleic acid construct encoding a multiepitope polypeptide, the multiepitope polypeptide comprising two or more polypeptides,
 wherein the two or more polypeptides include   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 8 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 having 8 or more consecutive amino acids from SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 8 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 8 or more consecutive amino acids from SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 5 or a fragment of SEQ ID NO: 5 having 8 or more consecutive amino acids from SEQ ID NO: 5, or   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 8 or more consecutive amino acids from SEQ ID NO: 6,   wherein, the two or more polypeptides are immunogenic to one or more influenza strains in a vertebrate expressing a major histocompatibility complex (MHC) allele, and wherein the two or more isolated polypeptides are not a complete influenza virus protein.   
     
     
         2 . The construct according to  claim 1 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 8 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 having 8 or more consecutive amino acids from SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 8 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 8 or more consecutive amino acids from SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 5 or a fragment of SEQ ID NO: 5 having 8 or more consecutive amino acids from SEQ ID NO: 5, and   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 8 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         3 . The construct according to  claim 1 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 8 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 8 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 8 or more consecutive amino acids from SEQ ID NO: 4, and   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 8 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         4 . The construct according to  claim 1 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 9 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 having 9 or more consecutive amino acids from SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 9 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 9 or more consecutive amino acids from SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 5 or a fragment of SEQ ID NO: 5 having 9 or more consecutive amino acids from SEQ ID NO: 5, or   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 9 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         5 . The construct according to  claim 4 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 9 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 having 9 or more consecutive amino acids from SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 9 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 9 or more consecutive amino acids from SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 5 or a fragment of SEQ ID NO: 5 having 9 or more consecutive amino acids from SEQ ID NO: 5, and   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 9 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         6 . The construct according to  claim 4 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 9 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 9 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 9 or more consecutive amino acids from SEQ ID NO: 4, and   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 9 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         7 . The construct according to  claim 1 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 5, or   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6.   
     
     
         8 . The construct according to  claim 7 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 5, and   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6.   
     
     
         9 . The construct according to  claim 7 , wherein the two or more polypeptides comprise
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4, and   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6.   
     
     
         10 . The composition according to  claim 1 , wherein the two or more polypeptides comprise
 a polypeptide comprising SEQ ID NO: 1,   a polypeptide comprising SEQ ID NO: 2,   a polypeptide comprising SEQ ID NO: 3,   a polypeptide comprising SEQ ID NO: 4,   a polypeptide comprising SEQ ID NO: 5, or   a polypeptide comprising SEQ ID NO: 6.   
     
     
         11 . The composition according to  claim 10 , wherein the two or more polypeptides comprise
 a polypeptide comprising SEQ ID NO: 1,   a polypeptide comprising SEQ ID NO: 2,   a polypeptide comprising SEQ ID NO: 3,   a polypeptide comprising SEQ ID NO: 4,   a polypeptide comprising SEQ ID NO: 5, and   a polypeptide comprising SEQ ID NO: 6.   
     
     
         12 . The composition according to  claim 10 , wherein the two or more polypeptides comprise
 a polypeptide comprising SEQ ID NO: 1,   a polypeptide comprising SEQ ID NO: 3,   a polypeptide comprising SEQ ID NO: 4, and   a polypeptide comprising SEQ ID NO: 6.   
     
     
         13 . The composition according to  claim 1 , wherein the one or more influenza strains includes an influenza A strain, an influenza B strain, or both an influenza A strain and an influenza B strain. 
     
     
         14 . A pharmaceutical composition comprising a recombinant nucleic acid construct of  claim 1  and an adjuvant. 
     
     
         15 . A recombinant nucleic acid construct encoding a polypeptide, the polypeptide being from 8 to 50 amino acids in length and selected from
 a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 8 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 having 8 or more consecutive amino acids from SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 8 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 8 or more consecutive amino acids from SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 5 or a fragment of SEQ ID NO: 5 having 8 or more consecutive amino acids from SEQ ID NO: 5, or   a polypeptide comprising an amino acid sequence with 85% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 8 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         16 . The construct according to  claim 15 , wherein the polypeptide comprises
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having 9 or more consecutive amino acids from SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 having 9 or more consecutive amino acids from SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3 or a fragment of SEQ ID NO: 3 having 9 or more consecutive amino acids from SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4 or a fragment of SEQ ID NO: 4 having 9 or more consecutive amino acids from SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 5 or a fragment of SEQ ID NO: 5 having 9 or more consecutive amino acids from SEQ ID NO: 5, or   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6 or a fragment of SEQ ID NO: 6 having 9 or more consecutive amino acids from SEQ ID NO: 6.   
     
     
         17 . The construct according to  claim 16 , wherein the polypeptide comprises
 a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 1,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 2,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 3,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 4,   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 5, or   a polypeptide comprising an amino acid sequence with 95% or more amino acid sequence identity to SEQ ID NO: 6.   
     
     
         18 . The composition according to  claim 17 , wherein the polypeptide comprises
 a polypeptide comprising SEQ ID NO: 1,   a polypeptide comprising SEQ ID NO: 2,   a polypeptide comprising SEQ ID NO: 3,   a polypeptide comprising SEQ ID NO: 4,   a polypeptide comprising SEQ ID NO: 5, or   a polypeptide comprising SEQ ID NO: 6.   
     
     
         19 . A pharmaceutical composition comprising a recombinant nucleic acid construct of  claim 15  and an adjuvant. 
     
     
         20 . The composition according to  claim 19 , which is immunogenic to one or more influenza virus strains.