Influenza Hemagglutinin And Neuraminidase Variants

Abstract

Polypeptides, polynucleotides, methods, compositions, and vaccines comprising influenza hemagglutinin and neuraminidase variants are provided.

1 . An isolated polypeptide comprising any one of the amino acid sequences SEQ ID NO:35-52 and 54-68. 
     
     
         2 . An immunogenic composition comprising an immunologically effective amount of the polypeptide of  claim 1 . 
     
     
         3 . An isolated polynucleotide encoding the polypeptide of  claim 1 . 
     
     
         4 . The polynucleotide of  claim 3 , wherein the polynucleotide is DNA. 
     
     
         5 . The polynucleotide of  claim 3 , wherein the polynucleotide is RNA. 
     
     
         6 . An immunogenic composition comprising an immunologically effective amount of the polynucleotide of  claim 3 . 
     
     
         7 . A reassortant influenza virus comprising the polynucleotide of  claim 3 . 
     
     
         8 . The virus of  claim 7 , wherein the virus comprises 6 internal genome segments from one or more donor viruses. 
     
     
         9 . The virus of  claim 8 , wherein one donor virus is A/Ann Arbor/6/60, or A/Puerto Rico/8/34. 
     
     
         10 . An immunogenic composition comprising an immunologically effective amount of the recombinant influenza virus of  claim 8 . 
     
     
         11 . A vector comprising the polynucleotide of  claim 3 . 
     
     
         12 . The vector of  claim 11 , wherein the vector is a plasmid, a cosmid, a phage, or a virus. 
     
     
         13 . The vector of  claim 11 , wherein the vector is an expression vector. 
     
     
         14 . An isolated cell comprising the vector of  claim 11 . 
     
     
         15 . The virus of  claim 8 , 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. 
     
     
         16 . A method for producing the reassortant influenza virus of  claim 7  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 any one of the amino acid sequences SEQ ID NO:35-52 and 54-68;   ii) culturing the population of host cells; and,   iii) recovering the influenza virus.   
     
     
         17 . The method of  claim 16 , 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. 
     
     
         18 . The influenza virus produced by the method of  claim 16 , wherein the influenza virus is suitable for administration in an intranasal vaccine formulation. 
     
     
         19 . The method of  claim 16 , wherein the first influenza strain is an influenza A strain. 
     
     
         20 . The method of  claim 16 , wherein the first influenza strain is A/Ann Arbor/6/60, or A/Puerto Rico/8/34. 
     
     
         21 . The method of  claim 16 , wherein the plurality of vectors is a plurality of plasmid vectors. 
     
     
         22 . The method of  claim 16 , wherein the population of host cells comprises one or more of: Vero cells, PerC6 cells, MDCK cells, 293T cells, or COS cells. 
     
     
         23 . The method of  claim 16 , wherein the method does not comprise use of a helper virus. 
     
     
         24 . The method of  claim 16 , wherein the plurality of vectors consists of eight vectors. 
     
     
         25 . An immunogenic composition comprising the polypeptide of  claim 1 . 
     
     
         26 . The composition of  claim 25 , further comprising an excipient. 
     
     
         27 . The composition of  claim 26 , wherein the excipient is a pharmaceutically acceptable excipient. 
     
     
         28 . An immunogenic composition comprising the polynucleotide of  claim 3 . 
     
     
         29 . The composition of  claim 28 , further comprising an excipient. 
     
     
         30 . The composition of  claim 29 , wherein the excipient is a pharmaceutically acceptable excipient. 
     
     
         31 . An immunogenic composition comprising the reassortant virus of  claim 7 . 
     
     
         32 . The composition of  claim 31  wherein the reassortant virus is a 6:2 reassortment virus comprising 6 internal genome segments from one or more donor viruses. 
     
     
         33 . The composition of  claim 32 , 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. 
     
     
         34 . The composition of  claim 33 , wherein one donor virus is A/Ann Arbor/6/60, or A/Puerto Rico/8/34. 
     
     
         35 . A live attenuated influenza vaccine comprising the composition of  claim 31 . 
     
     
         36 . The composition of  claim 31 , further comprising one or more pharmaceutically acceptable excipient. 
     
     
         37 . A method of prophylactic or therapeutic treatment of a viral infection in a subject, the method comprising: administering to the subject the virus of  claim 7  in an amount effective to produce an immunogenic response against the viral infection. 
     
     
         38 . The method of  claim 37 , wherein the subject is a mammal. 
     
     
         39 . The method of  claim 38 , wherein the mammal is a human. 
     
     
         40 . The method of  claim 37 , wherein the virus is formulated using at least one pharmaceutically acceptable excipient.