Genomic deletion in african swine fever vaccine allowing efficient growth in stable cell lines

Patent metadata

Field	Value
Publication number	US-2022193217-A1
Application number	US-202017130814-A
Country	US
Kind code	A1
Filing date	Dec 22, 2020
Priority date	Dec 22, 2020
Publication date	Jun 23, 2022
Grant date	—

Abstract

Provided herein are details on the construction of a recombinant African Swine Fever Virus (ASFV) live attenuated vaccine for prevention of ASF caused by various strains of ASFV, such as the highly virulent Georgia 2007 isolate (“ASFV-G”). An exemplary vaccine comprises a deletion of multiple genes allowing for industrial-scale growth in stable cell lines.

First claim

What is claimed is:

1 . A genetically modified virus, wherein the virus genome comprises a viral genome at least 95% identical to SEQ ID NO: 1.

2 . The virus of claim 1 , wherein the viral genome comprises SEQ ID NO:1.

3 . A vaccine composition against African Swine Fever Virus (ASFV), comprising the genetically modified virus of claim 1 .

4 . The vaccine composition of claim 3 , wherein the ASFV is ASFV-Georgia 2007 isolate (ASFV-G).

5 . A method for the protection of swine against ASFV, comprising administering to a swine a live attenuated vaccine comprising the genetically modified virus of claim 1 in an amount effective to protect said swine from clinical ASFV disease.

6 . The method of claim 5 , wherein the ASFV is ASFV-G.

7 . The method of claim 5 , wherein the amount effective to protect said swine from clinical ASFV disease is a vaccine comprising 10 2 -10 6 HAD 50 of the genetically modified virus of claim 1 .

8 . A recombinant ASFV mutant virus, a deletion, or partial deletion of each of ORFs MGF360-4L, MGF360-6L, X69R, MGF300-1L, MGF300-2R, MGF300-4L, MGF3608L, MGF360-9L, MGF360-10L, MGF360-11L.

9 . The recombinant virus of claim 8 , wherein the virus comprises a deletion of a genomic fragment at least 95% identical to SEQ ID NO:2.

10 . The recombinant virus of claim 8 , wherein the mutant ASFV is an ASFV-Georgia isolate.

11 . The recombinant virus of claim 8 , wherein the mutant ASFV comprises a genome at least 99% identical to SEQ ID NO: 1.

12 . A vaccine composition against ASFV-G, comprising the recombinant virus of claim 8 .

13 . A method for the protection of swine against ASFV, comprising administering to a swine a live attenuated vaccine comprising the recombinant virus of claim 8 in an amount effective to protect said swine from clinical ASFV disease.

14 . The method of claim 13 , wherein the ASFV is ASFV-G.

15 . The method of claim 14 , wherein the amount effective to protect said swine from clinical ASFV disease is a vaccine comprising 10 2 -10 6 HAD 50 of the genetically modified virus of claim 8 .

16 . A method of producing ASFV at titers of 10 4 -10 7 HAD 50 /mL in a cultured stable cell line, comprising inoculating the virus of claim 1 into said cultured stable cell line; incubating the inoculated cell line under conditions allowing for viral replication, and; growing said viruses to a titer of 10 4 -10 7 HAD 50 /mL.

17 . The method of claim 16 , wherein the stable cell line is a porcine fetal kidney cell line engineered to express bovine α V β 6 integrin.

18 . The method of claim 16 , wherein the ASFV virus genome comprises a viral genome at least 99% identical to SEQ ID NO: 1.

Assignees

• Us Agriculture

Inventors

• Gladue Douglas P
• Borca Manuel V

Classifications

• A61P31/20

  for DNA viruses · CPC title

• A61K2039/552

  Veterinary vaccine · CPC title

• A61K2039/5254

  avirulent or attenuated · CPC title

• A61K39/12 Primary

  Viral antigens · CPC title

• C12N2710/12021

  Viruses as such, e.g. new isolates, mutants or their genomic sequences · CPC title