Influenza hemagglutinin and neuraminidase variants

Abstract

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.