INFLUENZA HEMAGGLUTININ AND NEURAMINIDASE VARIANTS
Polypeptides, polynucleotides, methods, compositions, and vaccines comprising influenza hemagglutinin and neuraminidase variants are provided.
1 . A reassortant influenza virus comprising a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 39.
2 . The virus of claim 1 , wherein the virus comprises 6 internal genome segments from one or more donor viruses.
3 . The virus of claim 2 , wherein one donor virus is A/Ann Arbor/6/60, or A/Puerto Rico/8/34.
4 . An immunogenic composition comprising an immunologically effective amount of the recombinant influenza virus of claim 2 .
5 . The virus of claim 2 , wherein the 6 internal genome segments of the one or more donor viruses are selected for comprising 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 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 the amino acid sequence of SEQ ID NO: 39;
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 for comprising one or more phenotypic attributes selected from the group consisting of:
attenuated, cold-adapted and temperature-sensitive.
8 . An influenza virus produced by the method of claim 6 , wherein the influenza virus is suitable for administration in an intranasal vaccine formulation.
9 . The method of claim 6 , wherein the first influenza strain is an influenza A strain.
10 . The method of claim 6 , wherein the first influenza strain is A/Ann Arbor/6/60, or A/Puerto Rico/8/34.
11 . The method of claim 6 , wherein the plurality of vectors is a plurality of plasmid vectors.
12 . The method of claim 6 , wherein the population of host cells comprises one or more of: Vero cells, PerC6 cells, MDCK cells, 293T cells, or COS cells.
13 . The method of claim 6 , wherein the method does not comprise use of a helper virus.
14 . The method of claim 6 , wherein the plurality of vectors consists of eight vectors.
15 . An immunogenic composition comprising the reassortant virus of claim 1 .
16 . The composition of claim 15 wherein the reassortant virus is a 6:2 reassortant virus comprising 6 internal genome segments from one or more donor viruses.
17 . The composition of claim 16 , wherein the 6 internal genome segments of the one donor virus are selected for comprising one or more phenotypic attributes selected from the group consisting of: attenuated, cold adapted and temperature sensitive.
18 . The composition of claim 17 , wherein one donor virus is A/Ann Arbor/6/60, or A/Puerto Rico/8/34.
19 . A live attenuated influenza vaccine comprising the composition of claim 15 .
20 . The composition of claim 15 , further comprising one or more pharmaceutically acceptable excipients.
21 . A method of prophylactic or therapeutic treatment of an influenza A H3 infection in a subject, the method comprising: administering to the subject the virus of claim 1 in an amount effective to produce an immunogenic response against the viral infection.
22 . The virus of claim 1 , wherein the virus further comprises a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 40.
23 . The virus of claim 1 , wherein the virus is killed or inactivated.
24 . A method of prophylactic or therapeutic treatment of an influenza A H3 viral infection in a subject, the method comprising: administering to the subject the virus of claim 23 in an amount effective to produce an immunogenic response against the influenza A H3 viral infection.