Avian influenza vaccines and methods of making same
The disclosure relates to avian influenza vaccines produced through reverse genetics to protect from A/Turkey/Indiana/22-like virus, methods for producing such vaccines, and method for using such vaccines to protect poultry. The disclosure also relates to methods of producing influenza viruses comprising chimeric polynucleotides having an HA non-coding sequence from an apathogenic high growth influenza virus and a polynucleotide encoding an HA protein from a highly pathogenic avian influenza virus or variant thereof, and/or having a chimeric polynucleotide having an NA non-coding sequence from an apathogenic high growth influenza virus and a polynucleotide encoding an NA protein from a highly pathogenic avian influenza virus or variant thereof.
1 . A chimeric polynucleotide comprising:
(i) a hemagglutinin (HA) non-coding sequence from an A/Puerto Rico/8/34 (PR8) apathogenic high growth influenza virus and a polynucleotide encoding an attenuated HA protein from an A/turkey/Indian/22-003707-003/2022 highly pathogenic avian influenza virus or a variant thereof having at least 96% identity thereto, wherein the wild type A/turkey/Indian/22-003707-003/2022 HA nucleotide sequence is set forth in SEQ ID NO: 3; or
(ii) a neuraminidase (NA) non-coding sequence from an PR8 apathogenic high growth influenza virus and a polynucleotide encoding an NA protein from an A/turkey/Indian/22-003707-003/2022 highly pathogenic avian influenza virus or a variant thereof having at least 96% identity thereto, wherein the wild type A/turkey/Indian/22-003707-003/2022 NA nucleotide sequence is set forth in SEQ ID NO: 3; or
(iii) an NA non-coding sequence from an PR8 apathogenic high growth influenza virus and a polynucleotide encoding an NA protein from a A/Blue Winged Teal/Wyoming/AH0099021/2016 low pathogenic influenza virus or a variant thereof having at least 96% identity thereto, wherein the wild type A/Blue Winged Teal/Wyoming/AH0099021/2016 NA nucleotide sequence is set forth in SEQ ID NO: 3;
wherein the HA is attenuated by altering the highly pathogenic cleavage site into a low pathogenic cleavage site.
2 . A vector comprising a chimeric polynucleotide of claim 1 .
3 . The vector of claim 2 , wherein the vector is a plasmid, a cosmid, a phage, a virus, or a fragment of a virus.
4 . A cell comprising the vector of claim 2 .
5 . A rescued influenza virus comprising polynucleotides encoding M1/M2, NP, PB1, PA, PB2, and NS1/NS2 from an PR8 apathogenic high growth influenza A virus, a polynucleotide encoding an attenuated HA from a A/turkey/Indiana/22-003707-003/2022 high highly pathogenic avian influenza virus or a variant thereof having 96% identity thereto, and a polynucleotide encoding an N1, N2, N3, N4, N5, N6, N7, N8, or N9 NA; wherein the wild type A/turkey/Indian/22-003707-003/2022 HA nucleotide sequence is set forth in SEQ ID NO: 3;
wherein the HA is attenuated by altering the highly pathogenic cleavage site into a low pathogenic cleavage site.
6 . The rescued influenza virus from claim 5 , wherein the polynucleotide encoding NA is from A/turkey/Indiana/22-003707-003/2022 having the nucleotide sequence of SEQ ID NO: 4, A/Blue Winged Teal/Wyoming/AH0099021/2016 having the nucleotide sequence of SEQ ID NO: 5, or a variant thereof having 96% identity thereto.
7 . The rescued influenza virus of claim 6 , wherein the polynucleotide encoding NA is from A/turkey/Indiana/22-003707-003/2022 having the nucleotide sequence of SEQ ID NO: 4.
8 . The rescued influenza virus of claim 6 , wherein the polynucleotide encoding NA is from A/Blue Winged Teal/Wyoming/AH0099021/2016 having the nucleotide sequence of SEQ ID NO: 5.
9 . An expression vector comprising any one of the polynucleotides encoding M1/M2, NP, PB1, PA, PB2, NS1/NS2, HA, or NA of claim 5 .
10 . An immunogenic composition comprising an immunologically-effective amount of the rescued influenza virus of claim 5 .
11 . A method of producing an influenza virus, the method comprising introducing into a population of host cells a plurality of vectors comprising polynucleotides corresponding to at least six internal segments of an PR8 apathogenic high growth influenza virus, a chimeric polynucleotide comprising an HA non-coding sequence from an PR8 apathogenic high growth influenza virus and a polynucleotide encoding an attenuated HA protein from a A/turkey/Indiana/22-003707-003/2022 highly pathogenic avian influenza virus or a variant thereof having 96% identity thereto, and a chimeric polynucleotide comprising an NA non-coding sequence from an PR8 apathogenic high growth influenza virus and a polynucleotide encoding an N1, N2, N3, N4, N5, N6, N7, N8, or N9 NA protein or a variant thereof having 96% identity thereto, culturing the population of host cells at a temperature that is less or equal to about 35° C., and recovering the virus;
wherein the wild type A/turkey/Indian/22-003707-003/2022 HA nucleotide sequence is set forth in SEQ ID NO: 3; and
wherein the HA is attenuated by altering the highly pathogenic cleavage site into a low pathogenic cleavage site.
12 . The method of claim 11 , wherein the NA is from A/turkey/Indiana/22-003707-003/2022 having the nucleotide sequence of SEQ ID NO: 4, A/Blue Winged Teal/Wyoming/AH0099021/2016 having the nucleotide sequence of SEQ ID NO: 5, or a variant thereof having 96% identity thereto.
13 . The method of claim 12 , wherein the NA polynucleotide is from A/turkey/Indiana/22-003707-003/2022 having the nucleotide sequence of SEQ ID NO: 4, or A/Blue Winged Teal/Wyoming/AH0099021/2016 having the nucleotide sequence of SEQ ID NO: 5.