Influenza virus reassortment

Patent metadata

Field	Value
Publication number	US-10864264-B2
Application number	US-201916657082-A
Country	US
Kind code	B2
Filing date	Oct 18, 2019
Priority date	Mar 13, 2013
Publication date	Dec 15, 2020
Grant date	Dec 15, 2020

Abstract

New influenza donor strains for the production of reassortant influenza B viruses are provided.

First claim

The invention claimed is:

1. A method of preparing a reassortant influenza B virus comprising steps of:
(i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza B virus wherein the expression construct(s) encode the HA segment from a first influenza B virus and the NP and/or PB2 segment from a second influenza B virus which is a B/Victoria/2/87-like strain; and
(ii) culturing the culture host in order to produce a reassortant influenza B virus.

2. A method of preparing a reassortant influenza B virus comprising steps of:
(i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza B virus wherein the expression construct(s) encode the HA segment from a first influenza B virus and the NP segment from a second influenza B virus which is not B/Lee/40 or B/Ann Arbor/1/66 or B/Panama/45/90; and
(ii) culturing the culture host in order to produce a reassortant influenza B virus.

3. The method of claim 2 , wherein the NP and PB2 segments are from the second influenza B virus.

4. The method of claim 2 , wherein the second influenza B virus is a B/Victoria/2/87-like strain.

5. The method of claim 1 , wherein the PA, PB1, PB2, NP, NS and M segments are from the second influenza B virus.

6. The method of claim 1 , wherein the reassortant influenza B virus comprises backbone segments from two or more influenza B strains.

7. The method of claim 6 , wherein at least one backbone segment is from a B/Yamagata/16/88-like strain.

8. A method of preparing a reassortant influenza B virus comprising steps of
(i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza B virus comprising the HA segment from a B/Yamagata/16/88-like strain and at least one backbone segment from a B/Victoria/2/87-like strain; and
(ii) culturing the culture host in order to produce a reassortant influenza B virus.

9. The method of claim 8 , wherein two, three, four, five or six backbone segments are from the B/Victoria/2/87-like strain.

10. The method of claim 8 , wherein the ratio of segments from the B/Victoria/2/87-like strain and the B/Yamagata/16/88-like strain is 7:1, 6:2, 4:4, 3:5 or 1:7.

11. A method of preparing a reassortant influenza B virus comprising steps of
(i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza B virus comprising viral segments from a B/Victoria/2/87-like strain and a B/Yamagata/16/88-like strain, wherein the ratio of segments from the B/Victoria/2/87-like strain and the B/Yamagata/16/88-like strain is 1:7, 2:6, 3:5, 4:4, 5:3, 6:2 or 7:1; and
(ii) culturing the culture host in order to produce a reassortant influenza B virus.

12. The method of claim 10 , wherein the ratio is 7:1, 6:2, 4:4, 3:5 or 1:7.

13. The method of claim 1 , wherein the B/Victoria/2/87-like strain is B/Brisbane/60/08.

14. The method of claim 11 , wherein the B/Yamagata/16/88-like strain is B/Panama/45/90.

15. The method of claim 1 , wherein the NP segment encodes a protein which has at least 97%, at least 98%, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 4.

16. The method of claim 1 , wherein the PB2 segment encodes a protein which has at least 97%, at least 98%, at least 99% or 100% identity to the sequence of SEQ ID NO: 3.

17. The method of claim 1 , wherein the NS segment encodes a protein which has at least 97%, at least 98%, at least 99% or 100% identity with the sequence of SEQ ID NO: 35 and/or wherein the M1 segment encodes a protein which has at least 97%, at least 98%, at least 99% or 100% identity with the sequence of SEQ ID NO: 34.

18. The method of claim 1 , wherein the reassortant influenza B virus comprises:
a) a PA protein which has at least 97% identity, at least 98% identity, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 1; and/or
b) a PB1 protein which has at least 97% identity, at least 98% identity, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 2; and/or
c) a M1 protein which has at least 97% identity, at least 98%, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 5; and/or
d) a M2 protein which has at least 97% identity, at least 98% identity, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 6; and/or
e) a NS1 protein which has at least 97% identity, at least 98%, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 7; and/or
f) a NS2 protein which has at least 97% identity, at least 98%, at least 99% identity or 100% identity to the sequence of SEQ ID NO: 8.

19. A method of preparing a reassortant influenza B virus comprising steps of
(i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza B virus comprising:
a) the PA segment of SEQ ID NO: 11, the PB1 segment of SEQ ID NO: 12, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 16 and the M segment of SEQ ID NO: 15; or
b) the PA segment of SEQ ID NO: 11, the PB1 segment of SEQ ID NO: 31, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34; or
c) the PA segment of SEQ ID NO: 11, the PB1 segment of SEQ ID NO: 31, the PB2 segment of SEQ ID NO: 32, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 16 and the M segment of SEQ ID NO: 15; or
d) the PA segment of SEQ ID NO: 30, the PB1 segment of SEQ ID NO: 12, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 16 and the M segment of SEQ ID NO: 15, or
e) the PA segment of SEQ ID NO: 11, the PB1 segment of SEQ ID NO: 12, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34;
f) the PA segment of SEQ ID NO: 30, the PB1 segment of SEQ ID NO: 31, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 33, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34, or
g) the PA segment of SEQ ID NO: 30, the PB1 segment of SEQ ID NO: 31, the PB2 segment of SEQ ID NO: 32, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34, or
h) the PA segment of SEQ ID NO: 11, the PB1 segment of SEQ ID NO: 12, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 33, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34, or
i) the PA segment of SEQ ID NO: 11, the PB1 segment of SEQ ID NO: 12, the PB2 segment of SEQ ID NO: 32, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34, or
j) the PA segment of SEQ ID NO: 30, the PB1 segment of SEQ ID NO: 12, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34, or
k) the PA segment of SEQ ID NO: 30, the PB1 segment of SEQ ID NO: 31, the PB2 segment of SEQ ID NO: 13, the NP segment of SEQ ID NO: 14, the NS segment of SEQ ID NO: 35 and the M segment of SEQ ID NO: 34; and
(ii) culturing the culture host in order to produce a reassortant influenza B virus.

20. The method of claim 1 , further comprising the step (iii) of purifying the reassortant virus obtained in step (ii).

21. A reas

Assignees

• Seqirus Uk Ltd
• Synthetic Genomics Inc

Inventors

• Dormitzer Philip
• Mason Peter
• Suphaphiphat Pirada
• Gomila Raul

Classifications

• A61K39/12 Primary

  Viral antigens · CPC title

• C12N2760/16234

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N2760/16251

  Methods of production or purification of viral material · CPC title

• C12N2760/16252

  relating to complementing cells and packaging systems for producing virus or viral particles · CPC title

• C12N2760/16134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title