# US 8,784,838 B2 · H3 influenza A virus

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- Number: US 8,784,838 B2
- Document ID: US8784838B2
- Status: Granted
- Kind: B2
- Application number: 13/839,111
- Filed: 2013-03-15
- Published: 2013-08-15
- Granted: 2014-07-22
- Earliest claimed: 2005-01-11
- Assignees: Wisconsin Alumni Research Foundation
- Applicants: Wisconsin Alumni Research Foundation
- Inventors: Christopher W. Olsen; Gabriele A. Landolt; Alexander I. Karasin
- Web page: https://patentgrep.com/patent/US8784838B2

## CPC

- A61K 39/145: Orthomyxoviridae, e.g. influenza virus
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 9/0043: {Nose}
- A61K 39/12: Viral antigens
- C07K 14/005: from viruses
- C12N 1/00: Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- G01N 33/56983: {Viruses}
- A61K 2039/525: {Virus}
- A61K 2039/5252: {inactivated (killed)}
- A61K 2039/5254: {avirulent or attenuated}
- A61K 2039/54: {characterised by the route of administration}
- A61K 2039/543: {intranasal}
- A61K 2039/552: {Veterinary vaccine}
- A61K 2039/70: {Multivalent vaccine}
- C12N 2760/16121: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2760/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12R 2001/91: Cell lines {; Processes using cell lines}
- G01N 2333/11: Orthomyxoviridae, e.g. influenza virus
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The invention provides an isolated H3 equine influenza A virus, as well as methods of preparing and using the virus, and genes or proteins thereof.

## Claims

### Claim 1

An inactivated preparation of isolated H3 influenza virus comprising a gene segment with sequences for a HA having SEQ ID NO:1 or a HA having at least 99% amino acid sequence identity to SEQ ID NO:1 which HA does not have a valine at position 78 or an asparagine at position 159.

### Claim 2 (depends on 1)

The preparation of claim 1 wherein the virus comprises at least one of the following gene segments: a gene segment with sequences for a NA having SEQ ID NO:2 or having at least 95% amino acid sequence identity to SEQ ID NO:2, a gene segment with sequences for a PB1 having SEQ ID NO:3 or having at least 95% amino acid sequence identity to SEQ ID NO:3, a gene segment with sequences for a PB2 having SEQ ID NO:4 or having at least 95% amino acid sequence identity to SEQ ID NO:4, a gene segment with sequences for a PA having SEQ ID NO:5 or having at least 95% amino acid sequence identity to SEQ ID NO:5, a gene segment with sequences for a NP having SEQ ID NO:6 or having at least 95% amino acid sequence identity to SEQ ID NO:6, a gene segment with sequences for a M1 having SEQ ID NO:7 or having at least 95% amino acid sequence identity to SEQ ID NO:7, a gene segment with sequences for a M2 having SEQ ID NO:17 or having at least 95% amino acid sequence identity to SEQ ID NO:17, a gene segment with sequences for a NS1 having SEQ ID NO:8 or having at least 95% amino acid sequence identity to SEQ ID NO:8, or a gene segment with sequences for a NS2 having SEQ ID NO:18 or having at least 95% amino acid sequence identity to SEQ ID NO:18.

### Claim 3 (depends on 1)

The preparation of claim 1 wherein the virus comprises negative-strand nucleic acid corresponding to nucleic acid sequences encoding at least one of SEQ ID NOs:1-8, 17 or 18.

### Claim 4 (depends on 1)

The preparation of claim 1 wherein the virus has HA having SEQ ID NO:1 and NA having SEQ ID NO:2.

### Claim 5 (depends on 1)

A method to immunize a mammal against influenza, comprising administering to the mammal an effective amount of the preparation of claim 1.

### Claim 6 (depends on 5)

The method of claim 5 wherein the mammal is a dog or a horse.

### Claim 7 (depends on 1)

A vaccine comprising the preparation of claim 1 in an amount effective to induce a prophylactic or therapeutic response against influenza infection.

### Claim 8 (depends on 7)

The vaccine of claim 7 further comprising a different isolated influenza virus.

### Claim 9 (depends on 7)

The vaccine of claim 7 wherein the isolated influenza virus is a reassortant virus.

### Claim 10 (depends on 7)

The vaccine of claim 7 further comprising an adjuvant.

### Claim 11 (depends on 7)

The vaccine of claim 7 further comprising a pharmaceutically acceptable carrier.

### Claim 12 (depends on 11)

The vaccine of claim 11 wherein the carrier is suitable for intramuscular administration.

### Claim 13 (depends on 7)

The vaccine of claim 7 which is in freeze-dried form.

### Claim 14

An isolated H3 influenza virus comprising a HA1 protein encoded by a gene segment with sequences for a HA having SEQ ID NO:1 or a HA having at least 95% amino acid sequence identity to SEQ ID NO:1 which HA has an alanine at position 78 and a serine at position 159.

### Claim 15 (depends on 14)

A vaccine comprising the virus of claim 14 in an amount effective to induce a prophylactic or therapeutic response against influenza infection.

### Claim 16 (depends on 15)

The vaccine of claim 15 further comprising a different isolated influenza virus.

### Claim 17 (depends on 15)

The vaccine of claim 15 wherein the isolated influenza virus is an attenuated virus.

### Claim 18 (depends on 15)

The vaccine of claim 15 wherein the isolated influenza virus is a reassortant virus.

### Claim 19 (depends on 15)

The vaccine of claim 15 wherein the influenza virus has been altered by chemical, physical or molecular means.

### Claim 20 (depends on 15)

The vaccine of claim 15 further comprising an adjuvant.

### Claim 21 (depends on 15)

The vaccine of claim 15 further comprising a pharmaceutically acceptable carrier.

### Claim 22 (depends on 21)

The vaccine of claim 21 wherein the carrier is suitable for intranasal or intramuscular administration.

### Claim 23 (depends on 14)

The vaccine of claim 14 which is in freeze-dried form.

### Claim 24 (depends on 14)

A method to immunize a mammal against influenza, comprising administering to the mammal an effective amount of the virus of claim 14.

### Claim 25 (depends on 24)

The method of claim 24 wherein the mammal is a dog or a horse.

### Claim 26 (depends on 14)

A diagnostic method comprising contacting a physiological sample of an animal suspected of containing anti-influenza virus antibodies with the virus of claim 14, and determining whether the sample comprises antibodies specific for the virus.