Swine influenza A virus vaccine comprising a nucleic acid construct having a specific order of genes
The present invention relates to a nucleic acid construct that encodes, in this order, a first Swine influenza A (IAV-S) hemagglutinin (HA) antigen of the Scot/94 lineage and a second Swine influenza A (IAV-S) hemagglutinin (HA) antigen of the Eurasian avian-like (EA) lineage, and a nucleic acid construct that encodes, in this order, a first IAV-S HA antigen of the Gent/84 lineage and a second IAV-S HA antigen of pandemic09 (pdm09) lineage. In other embodiments, the present invention relates to RNA replicon particles comprising one or both nucleic acid constructs, an immunogenic composition, such as a vaccine, which may be used against influenza A virus infections, and comprising the replicon particles. Further provided are methods of making the vaccine and use of the vaccine.
1 . A nucleic acid construct comprising, in the order from 5′ to 3′ of its nucleic acid sequence:
a first nucleic acid sequence encoding a first HA antigen of a Swine influenza A virus (IAV-S) of the A/swine/Scotland/410440/1994-like H1 hu N2 (Scot/94) lineage, and
a second nucleic acid sequence encoding a second HA antigen of IAV-S of the Eurasian avian-like H1 av N1 (EA) lineage.
2 . The nucleic acid construct of claim 1 , wherein the first HA antigen is from strain A/swine/Italy/3033-1/2015 (H1N2).
3 . The nucleic acid construct of claim 2 , wherein the first HA antigen encoded by the first 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.
4 . The nucleic acid construct of claim 1 , wherein the second HA antigen is from strain A/swine/Italy/28762-3/2013 (H1N1).
5 . The nucleic acid construct of claim 4 , wherein the second HA antigen encoded by the second 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.
6 . A nucleic acid construct comprising, in the order from 5′ to 3′ of its nucleic acid sequence:
a first nucleic acid sequence encoding a first HA antigen of IAV-S of the A/swine/Gent/1/1984-like H3N2 (Gent/84) lineage, and
a second nucleic acid sequence encoding a second HA antigen of IAV-S of the A(H1N1)pdm09 (pdm09) lineage.
7 . The nucleic acid construct of claim 6 , wherein the first HA antigen is from strain A/swine/Italy/240849/2015 (H3N2).
8 . The nucleic acid construct of claim 7 , 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.
9 . The nucleic acid construct of claim 7 , wherein the second HA antigen is from strain A/swine/England/373/2010 (H1N1).
10 . The nucleic acid construct of claim 9 , 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 sequence having at least 95% sequence identity thereof.
11 . An RNA replicon particle comprising the nucleic acid construct of claim 1 .
12 . An RNA replicon particle comprising the nucleic acid construct of claim 6 .
13 . The RNA replicon particle of claim 11 , which is an alphavirus RNA replicon particle.
14 . The RNA replicon particle of claim 13 , which is a Venezuelan Equine Encephalitis (VEE) alphavirus RNA replicon particle.
15 . An immunogenic composition, comprising the alphavirus RNA replicon particle of claim 14 .
16 . A vaccine comprising the immunogenic composition of claim 15 .
17 . The vaccine of claim 16 , which is a nonadjuvanted vaccine.
18 . The vaccine of claim 16 , 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.
19 . 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 16 .