Influenza hemagglutinin and neuraminidase variants
Polypeptides, polynucleotides, methods, compositions, and vaccines comprising influenza hemagglutinin and neuraminidase variants are provided.
1. A reassortant influenza a virus comprising a polynucleotide encoding a polypeptide comprising the amino acid sequence SEQ ID NO: 43.
2. The reassortant influenza a virus of claim 1 , wherein the virus comprises 6 internal genome segments from one or more donor viruses.
3. The reassortant influenza a virus of claim 2 , wherein one or more donor viruses are A/Ann Arbor/6/60, or A/Puerto Rico/8/34.
4. An immunogenic composition comprising an immunologically effective amount of the reassortant influenza a virus of claim 2 .
5. The reassortant influenza a virus of claim 2 , wherein the 6 internal genome segments of the one or more donor viruses are selected as conferring one or more phenotypic attributes selected from the group consisting of: attenuated, cold-adapted and temperature-sensitive.
6. A method for producing the reassortant influenza a virus of claim 1 in cell culture, the method comprising:
i) introducing a plurality of vectors into a population of host cells capable of supporting replication of influenza viruses, which plurality of vectors comprises nucleotide sequences corresponding to at least 6 internal genome segments of a first influenza strain; and one genome segment encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 43;
ii) culturing the population of host cells; and
iii) recovering the influenza virus.
7. The method of claim 6 , wherein the at least 6 internal genome segments of the first influenza virus strain are selected as conferring one or more phenotypic attributes selected from the group consisting of: attenuated, cold-adapted and temperature-sensitive.
8. A reassortant influenza a virus produced according to a method comprising:
i) introducing a plurality of vectors into a population of host cells capable of supporting replication of influenza viruses, which plurality of vectors comprises nucleotide sequences corresponding to at least 6 internal genome segments of a first influenza strain; and one genome segment encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 43;
ii) culturing the population of host cells; and
iii) recovering reassortant influenza a virus, wherein the reassortant influenza virus recovered is suitable for administration in an intranasal vaccine formulation.
9. The method of claim 6 , wherein the plurality of vectors consists of eight vectors.
10. An immunogenic composition comprising the reassortant influenza a virus of claim 1 .
11. The immunogenic composition of claim 10 , wherein the reassortant influenza a virus is a 6:2 reassortant virus comprising 6 internal genome segments from one or more donor viruses.
12. The immunogenic composition of claim 11 , wherein the 6 internal genome segments of the one donor virus are selected as conferring one or more phenotypic attributes selected from the group consisting of: attenuated, cold adapted and temperature sensitive.
13. The immunogenic composition of claim 12 , wherein the one or more donor viruses are A/Ann Arbor/6/60, or A/Puerto Rico/8/34.
14. A live attenuated influenza a vaccine comprising the composition of claim 10 .
15. The immunogenic composition of claim 10 , further comprising one or more pharmaceutically acceptable excipients.
16. A method of inducing an immunogenic response against an influenza a viral infection in a subject, the method comprising: administering to the subject the reassortant influenza a virus of claim 1 in an amount effective to produce an immunogenic response against the viral infection.
17. The reassortant influenza a virus of claim 1 , wherein the virus further comprises a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 44.
18. The reassortant influenza a virus of claim 1 , wherein the virus is killed or inactivated.
19. A method of inducing an immunogenic response against an influenza a viral infection in a subject, the method comprising: administering to the subject the reassortant influenza a virus of claim 18 in an amount effective to produce an immunogenic response against the influenza viral infection.
20. The method of claim 6 , wherein the virus further comprises a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 44.