Infectious hepatitis E virus genotype 3 recombinants

Patent metadata

Field	Value
Publication number	US-9850468-B2
Application number	US-201514918466-A
Country	US
Kind code	B2
Filing date	Oct 20, 2015
Priority date	Jan 10, 2011
Publication date	Dec 26, 2017
Grant date	Dec 26, 2017

Abstract

The invention relates to the discovery of an HEV strain from a chronically infected patient. The virus grow unusually well in numerous cell cultures. Thus, the invention provides cell cultures, vectors, and vaccine compositions based on the virus.

First claim

What is claimed is:

1. An infectious hepatitis E virus (HEV) type 3 cDNA clone, wherein the infectious clone comprises an insert, relative to the HEV nucleic acid sequence set forth in SEQ ID NO:5, in the region of the nucleic acid sequence that encodes the hypervariable region of ORF1, wherein the insert encodes a polypeptide of 20 to 100 amino acids in length having at least 85% identity to a human ribosomal S17 or S19 protein.

2. The infectious HEV type 3 cDNA clone of claim 1 , wherein the insert encodes a polypeptide of 20 to 100 amino acids in length of the human ribosomal S17 or S19 protein.

3. The infectious HEV type 3 cDNA clone of claim 1 , wherein the insert encodes a polypeptide sequence that comprises a sequence of at least 30 amino acids in length that has at least 85% identity over its length to SEQ ID NO:9.

4. The infectious HEV type 3 cDNA clone of claim 1 , wherein the insert is positioned in the hypervariable region of ORF1 such that the first amino acid encoded by the insert occurs at position 750 relative to SEQ ID NO:6.

5. An infectious hepatitis E virus (HEV) cDNA clone, wherein the cDNA clone has at least 75% sequence identity to SEQ ID NO:1 and comprises an insert in the region of the nucleic acid sequence that encodes the hypervariable region of ORF1, relative to the HEV nucleic acid sequence set forth in SEQ ID NO:5, wherein the insert comprises a region of 20 to 100 amino acids in length that has at least 85% identity to a human ribosomal S17 or S19 protein.

6. The HEV cDNA clone of claim 5 , wherein the insert encodes a polypeptide of 20 to 100 amino acids in length of the human ribosomal S17 or S19 protein.

7. The HEV cDNA clone of claim 6 , wherein the cDNA clone has at least 85% identity to SEQ ID NO:1.

8. The HEV cDNA clone of claim 5 , wherein the insert encodes an in-frame amino acid sequence of at least 30 amino acids in length that has at least 85% identity over its length to SEQ ID NO:9.

9. An infectious hepatitis E virus genotype 1 cDNA clone comprising a hepatitis E virus (HEV) nucleic acid sequence, wherein the infectious clone comprises an insert in the region of the nucleic acid sequence that encodes the hypervariable region of ORF1, wherein the insert encodes a polypeptide of 20 to 100 amino acids in length having at least 85% identity to a human ribosomal S17 or S19 protein.

10. The cDNA clone of claim 9 , wherein the insert encodes a polypeptide of 20 to 100 amino acids in length of the human ribosomal S17 or S19 protein.

11. The cDNA clone of claim 10 , wherein the insert encodes an in-frame polypeptide sequence of at least 30 amino acids in length that has at least 85% identity over its length to SEQ ID NO:9.

12. A cell culture system comprising cells that comprise a cDNA clone of claim 1 .

13. A method of producing an immunogenic composition, the method comprising introducing an RNA obtained from a cDNA clone of claim 1 into cells and obtaining virus produced by the RNA.

14. The method of claim 13 , wherein the RNA is introduced into the cells as virus particles.

15. The method of claim 13 , wherein the cells are MRC 5 lung cells.

16. A method of assessing the virus status of a product of interest subjected to a virus treatment process, the method comprising:
adding HEV virus produced using a cDNA clone of claim 1 into the product;
subjecting the product to a virus treatment process that removes and/or inactivates virus present in the product; and
determining the level of added virus in the product remaining after or during the virus treatment process.

17. The method of claim 16 , wherein the product of interest is water, blood, or food for animal consumption.

18. The method of claim 17 , wherein the food is for human consumption.

Assignees

• Us Health
• Royal Cornwall Hospital Trust

Inventors

• Emerson Suzanne U
• Shukla Priyanka
• Nguyen Hanh T
• Purcell Robert H
• Dalton Harry R
• Bendall Richard

Classifications

• A61P31/14

  for RNA viruses · CPC title

• C12N2770/28121

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

• C12N2770/28122

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

• C12Q1/707

  non-A, non-B Hepatitis, excluding hepatitis D · CPC title

• C12N15/86

  Viral vectors · CPC title