Polyvalent influenza virus-like particle (vlp) compositions

Abstract

Polyvalent influenza virus-like particles (VLPs) comprising influenza antigenic polypeptides are described. Also described are compositions comprising these polyvalent VLPs as well as methods of making and using these VLPs.

What is claimed is: 
     
         1 . A noninfectious polyvalent influenza VLP comprising an influenza M1 protein and three or more antigenic influenza glycoproteins, wherein
 (i) the polyvalent VLP comprises two or more antigenic hemagglutinin (HA) glycoproteins-derived from two or more different Type A strains influenza strains and one or more antigenic hemagglutinin (HA) glycoproteins-derived from a Type B strain;   (ii) the polyvalent VLP comprises less than all of the proteins of an influenza polymerase complex;   (iii) the three or more antigenic glycoproteins are displayed on the surface of the VLP.   
     
     
         2 . The VLP of  claim 1 , further comprising an M2 protein. 
     
     
         3 . The VLP of  claim 2 , wherein the M2 protein comprises an amino acid modification as compared to a wild-type M2 protein. 
     
     
         4 . The VLP of  claim 1 , further comprising an influenza nucleoprotein (NP). 
     
     
         5 . The VLP of  claim 1 , wherein the VLP further comprises one or more NA glycoproteins. 
     
     
         6 . The VLP of  claim 1 , wherein the M1 protein comprises a nuclear localization signal and further wherein the nuclear localization signal (NLS) is modified by deletion, insertion or substitution of one or more residues. 
     
     
         7 . The VLP of  claim 1 , wherein the M1 protein comprises one or more amino acids involved in protein-protein interactions and further wherein one or more amino acid residues involved in protein-protein interactions are modified. 
     
     
         8 . The VLP of  claim 1 , wherein one or more L-domains are introduced into the M1 protein. 
     
     
         9 . The VLP of  claim 1 , comprising a fusion of M1 and an immunomodulatory polypeptide. 
     
     
         10 . The VLP of  claim 1 , wherein at least one of the antigenic glycoproteins is chimeric, wherein the transmembrane domain of the at least one antigen glycoprotein is replaced with a transmembrane domain from a different influenza stain. 
     
     
         11 . The VLP of  claim 1 , wherein at least one of the antigenic glycoproteins is chimeric, wherein the cytoplasmic tail region of the glycoprotein is replaced with a cytoplasmic tail region from a different influenza stain. 
     
     
         12 . An isolated host cell comprising a polyvalent VLP according to  claim 1 . 
     
     
         13 . An isolated packaging cell for producing a polyvalent VLP according to  claim 1 , the packaging cell stably or transiently transfected with one or more influenza protein-encoding sequences such that the polyvalent VLP is produced by the cell. 
     
     
         14 . The isolated packaging cell line of  claim 13 , wherein a polynucleotide encoding M1 is stably transfected into the cell. 
     
     
         15 . The isolated packaging cell line of  claim 13 , wherein the cell is a mammalian cell line. 
     
     
         16 . An immunogenic composition comprising the polyvalent VLP of  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         17 . The immunogenic composition of  claim 16 , further comprising an adjuvant. 
     
     
         18 . A method of producing a VLP according to  claim 1 , the method comprising the steps of:
 expressing one or more polynucleotides encoding the M1 and at least three influenza glycoproteins of the VLP in a suitable host cell under conditions such that the VLPs assemble in the host cell;   isolating the assembled VLPs from the host cell.   
     
     
         19 . The method of  claim 18 , wherein the host cell is selected from the group consisting of a mammalian cell, an insect cell, a yeast cell, a plant cell and a fungal cell. 
     
     
         20 . The method of  claim 18 , wherein the one or more polynucleotides are expressed from an expression vector. 
     
     
         21 . The method of  claim 20 , wherein the polynucleotides are operably linked to control elements compatible with expression in the selected host cell. 
     
     
         22 . The method of  claim 20 , wherein the expression vector is selected from the group consisting of a plasmid, a viral vector, a baculovirus vector and a non-viral vector. 
     
     
         23 . The method of  claim 18 , wherein one or more of the polynucleotides are stably integrated into the host cell. 
     
     
         24 . A method of generating an immune response in a subject to three or more influenza viruses, the method comprising the step of administering a composition comprising a polyvalent VLP according to  claim 1  to the subject. 
     
     
         25 . The method of  claim 24 , wherein the composition is administered intranasally. 
     
     
         26 . The method of  claim 24 , wherein the composition is administered in a multiple dose schedule.