Swine influenza A virus vaccine comprising two distinct RNA replicon particles
The present invention relates to an immunogenic composition comprising first and second RNA replicon particles. The first RNA replicon particle comprises a nucleic acid construct comprising first and second nucleic acid sequences encoding first and second hemagglutinin (HA) antigens of a Swine influenza A virus (IAV-S). The first HA antigen is a of the A/swine/Gent/1/1984-like H3N2 (Gent/84) lineage, and the second HA antigen is of the A(H1N1)pdm09 (pdm09) lineage. The second RNA replicon particle comprises a nucleic acid construct comprising third and fourth nucleic acid sequences encoding third and fourth HA antigens of S. The third HA antigen is of the A/swine/Scotland/410440/1994-like H1 hu N2 (Scot/94) lineage, and the fourth HA antigen is of the Eurasian avian-like H1 av N1 (EA) lineage. In other embodiments, the present invention relates to a vaccine, which may be used against influenza A virus infection, and comprising the immunogenic composition. Further provided are methods of making the vaccine and use of the vaccine.
1 . An immunogenic composition comprising a first and a second RNA replicon particle,
wherein the first RNA replicon particle is an alphavirus RNA replicon particle and comprises a nucleic acid construct comprising a first nucleic acid sequence encoding a first hemagglutinin (HA) antigen of a Swine influenza A virus (IAV-S) and a second nucleic acid sequence encoding a second HA antigen of a IAV-S, wherein
the first HA antigen is of the A/swine/Gent/1/1984-like H3N2 (Gent/84) lineage, and
the second HA antigen is of the A (H1N1)pdm09 (pdm09) lineage,
wherein the second RNA replicon particle is an alphavirus RNA replicon particle and comprises a nucleic acid construct comprising a third nucleic acid sequence encoding a third HA antigen of IAV-S and a fourth nucleic acid sequence encoding a fourth HA antigen of IAV-S, wherein
the third HA antigen is of the A/swine/Scotland/410440/1994-like H1 hu N2 (Scot/94) lineage, and
the fourth HA antigen is of the Eurasian avian-like H1 av N1 (EA) lineage.
2 . The immunogenic composition of claim 1 , wherein the first HA antigen of the Gent/84 lineage of the first RNA replicon particle is from strain A/swine/Italy/240849/2015 (H3N2).
3 . The immunogenic composition of claim 1 , wherein the first HA antigen encoded by the first nucleic acid sequence comprises an amino acid sequence of SEQ ID NO: 9 or an amino acid sequence having at least 90% sequence identity thereof.
4 . The immunogenic composition of claim 1 , wherein the second HA antigen of the pdm lineage of the first RNA replicon particle is from strain A/swine/England/373/2010 (H1N1).
5 . The immunogenic composition of claim 1 , wherein the second HA antigen encoded by the second nucleic acid sequence comprises an amino acid sequence of SEQ ID NO: 12 or an amino acid having at least 95% sequence identity thereof.
6 . The immunogenic composition of claim 1 , wherein the third HA antigen of the Scot94 lineage of the second RNA replicon particle is from strain A/swine/Italy/3033-1/2015 (H1N2).
7 . The immunogenic composition of claim 1 , wherein the third HA antigen encoded by the third nucleic acid sequence comprises an amino acid sequence of SEQ ID NO: 3 or an amino acid sequence having at least 85% sequence identity thereof.
8 . The immunogenic composition of claim 1 , wherein the fourth HA antigen of the EA lineage of the second RNA replicon particle is from strain A/swine/Italy/28762-3/2013 (H1N1).
9 . The immunogenic composition of claim 1 , wherein the fourth HA antigen encoded by the fourth nucleic acid sequence comprises an amino acid sequence of SEQ ID NO: 6 or an amino acid sequence having at least 90% sequence identity thereof.
10 . The immunogenic composition of claim 1 , which is adapted for simultaneous administration of the first and second RNA replicon particles.
11 . The immunogenic composition of claim 1 , wherein the alphavirus RNA replicon particle is a Venezuelan Equine Encephalitis Virus (VEEV) alphavirus RNA replicon particle.
12 . A vaccine comprising the immunogenic composition of claim 11 .
13 . The vaccine of claim 12 , which is a nonadjuvanted vaccine.
14 . The vaccine of claim 12 , which comprises an adjuvant selected from the group consisting of a biodegradable oil, an oil-in-water emulsion with 2.5-50% (v/v) mineral oil, and a biodegradable oil mixed with an oil-in-water emulsion with 2.5-50% (v/v) mineral oil.
15 . A method of immunizing a porcine against a swine influenza A virus, the method comprising administering to the porcine an immunologically effective amount of the vaccine of claim 13 .
16 . A method of immunizing a porcine against a swine influenza A virus, the method comprising administering to the porcine an immunologically effective amount of the vaccine of claim 12 .
17 . A method of immunizing a porcine against a swine influenza A virus, the method comprising administering to the porcine an immunologically effective amount of the vaccine of claim 13 .
18 . A vaccine comprising the immunogenic composition of claim 1 .