Attenuated influenza vaccines and uses thereof

Patent metadata

Field	Value
Publication number	US-9878032-B2
Application number	US-201414905320-A
Country	US
Kind code	B2
Filing date	Jul 18, 2014
Priority date	Jul 19, 2013
Publication date	Jan 30, 2018
Grant date	Jan 30, 2018

Abstract

Provided herein are attenuated influenza viruses and methods of making attenuated influenza viruses.

First claim

What is claimed is:

1. A modified influenza A virus comprising a PB1 polymerase, wherein the PB1 polymerase comprises a leucine to glutamine substitution (L319Q) at an amino acid corresponding to amino acid 319 of SEQ ID NO: 2.

2. The virus of claim 1 , wherein the PB1 polymerase further comprises one or more mutations selected from the group consisting of a lysine to glutamic acid substitution at position 391 (K391E), a glutamic acid to glycine substitution at position 581 (E581G) and an alanine to threonine substitution at position 661 (A661T).

3. The virus of claim 1 , wherein the virus further comprises a PB2 polymerase comprising an asparagine to serine substitution at position 265 (N265S).

4. The virus of claim 1 , wherein the virus further comprises an influenza virus nucleoprotein (NP) comprising an aspartic acid to glycine substitution at position 35 (D35G).

5. The virus of claim 1 , wherein the influenza A virus is selected from the group consisting of an H2N2 virus, an H3N2 virus, an H1N1 virus, an H9N2 virus and an H5N1 virus.

6. The virus of claim 1 , wherein the influenza A virus is A/Ann Arbor/6/60 (H2N2).

7. The virus of claim 1 , wherein the virus is a live attenuated influenza A virus with reduced growth from about 37° C. to about 39° C., as compared to an influenza A virus comprising a PB1 polymerase lacking a leucine to glutamine substitution (L319Q) at an amino acid corresponding to amino acid 319 of SEQ ID NO: 2.

8. An immunogenic composition comprising the virus of claim 1 and a pharmaceutically acceptable carrier.

9. A method for eliciting an immune response against an influenza A virus in a subject, comprising administering an effective dose of the immunogenic composition of claim 8 to the subject.

10. A method of producing an influenza A virus, comprising:
a) transfecting a population of host cells with one or more vectors comprising
i) nucleic acid sequences encoding the internal genome segments of an influenza A virus and;
ii) a nucleic acid encoding a PB1 polymerase with a leucine to glutamine substitution (L319Q) at an amino acid corresponding to amino acid 319 of SEQ ID NO: 2;

b) culturing the host cells; and
c) recovering the modified influenza A virus.

11. The method of claim 10 , wherein the nucleic acid encoding the PB1 polymerase further comprises one or more mutations selected from the group consisting of a lysine to glutamic acid substitution at position 391 (K391E), a glutamic acid to glycine substitution at position 581 (E581G) and an alanine to threonine substitution at position 661 (A661T).

12. The method of claim 10 , further comprising transforming the cells with a nucleic acid encoding a PB2 polymerase comprising a N265S mutation.

13. The method of claim 10 , further comprising transforming the cells with a nucleic acid encoding an NP protein comprising a D35G mutation.

14. The method of claim 10 , wherein the influenza A virus is selected from the group consisting of an H2N2 virus, an H3N2 virus, an H1N1 virus, an H9N2 virus and an H5N1 virus.

15. The method of claim 10 , wherein the cells are Vero cells, MDCK cells or CEK cells.

16. A method for producing an influenza A virus comprising:
a) infecting a population of cells with the virus of claim 1 ;
b) culturing the cells;
c) harvesting the virus from the culture of step b).

17. The method of claim 16 , wherein the cell is a mammalian cell or an avian cell.

18. A population of isolated cells comprising the virus of claim 1 .

Assignees

• Univ Rochester

Inventors

• Cox Andrew
• Dewhurst Stephen
• Treanor John
• Kim Baek

Classifications

• C12N2760/16122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C12N2760/00062

  by genetic engineering · CPC title

• C12N2760/00051

  Methods of production or purification of viral material · CPC title

• C12N7/04

  Inactivation or attenuation; Producing viral sub-units · CPC title

• A61K39/145 Primary

  Orthomyxoviridae, e.g. influenza virus · CPC title