Genetically stable recombinant modified vaccinia ankara (rMVA) vaccines and methods of preparation thereof

Patent metadata

Field	Value
Publication number	US-10603375-B2
Application number	US-201715589857-A
Country	US
Kind code	B2
Filing date	May 8, 2017
Priority date	Jun 5, 2009
Publication date	Mar 31, 2020
Grant date	Mar 31, 2020

Abstract

A method of preventing, controlling or treating CMV infections with a vaccine comprising an immunologically effective amount of a fusion cytomegalovirus (CMV) protein antigen comprising a nucleotide sequence encoding two or more antigenic portions of Immediate-Early Gene-1 or Immediate-Early Gene-2 (IEfusion), wherein the antigenic portions elicit an immune response when expressed by a vaccine.

First claim

What is claimed is:

1. A method of controlling or treating a cytomegalovirus (CMV) infection in a subject comprising:
administering to a subject in need thereof, a composition comprising an immunologically effective amount of a recombinant modified vaccinia Ankara (rMVA) virus, wherein the rMVA virus comprises a fusion nucleotide sequence which comprises SEQ ID NO: 11, and which encodes an IEfusion CMV protein antigen, said fusion nucleotide sequence comprising a nucleotide sequence encoding an Immediate-Early Gene-1 (IE1) antigenic portion directly fused to a nucleotide sequence encoding an Immediate-Early Gene-2 (IE2) antigenic portion, wherein
(i) the nucleotide sequence encoding the IE1 antigenic portion includes a nucleotide sequence encoding IE1 exon 4 (IE1/e4);
(ii) the nucleotide sequence encoding the IE2 antigenic portion is a nucleotide sequence encoding IE2 exon 5 (IE2/e5); or
(iii) both (i) and (ii), thereby controlling or treating the CMV infection in the subject.

2. The method of claim 1 , wherein the composition further comprises a nucleotide sequence which encodes at least one CMV antigen or a combination of antigens selected from the group consisting of HCMV-pp65, and glycoprotein B (gB).

3. The method of claim 1 , wherein the rMVA virus is genetically stable and maintains immunogenicity after serial passage, and wherein the DNA sequence of the IE1 or IE2 gene and the expression of the IE1 or IE2 gene is substantially unchanged over the time of serial passage.

4. A method of controlling or treating a cytomegalovirus (CMV) infection in a subject comprising:
administering to a subject in need thereof, a composition comprising: an immunologically effective amount of a recombinant modified vaccinia Ankara (rMVA) virus, wherein the rMVA virus comprises a fusion nucleotide sequence, which comprises SEQ ID NO: 11, and which encodes an IEfusion CMV protein antigen, said fusion nucleotide sequence comprising a nucleotide sequence encoding an Immediate-Early Gene-1 (IE1) antigenic portion directly fused to a nucleotide sequence encoding an Immediate-Early Gene-2 (IE2) antigenic portion, wherein
(i) the nucleotide sequence encoding the IE1 antigenic portion includes a nucleotide sequence encoding IE1 exon 4 (IE1/e4);
(ii) the nucleotide sequence encoding the IE2 antigenic portion is a nucleotide sequence encoding IE2 exon 5 (IE2/e5); or
(iii) both (i) and (ii), thereby controlling or treating the CMV infection in the subject,

wherein the composition is
produced by:
a) constructing a transfer plasmid vector comprising a modified H5 (mH5) promoter operably linked to a DNA sequence encoding the IEfusion CMV protein antigen, wherein the expression of said DNA sequence is under the control of the mH5 promoter;
b) generating the rMVA virus by transfecting one or more plasmid vectors obtained from step a) into cells infected with wild type MVA;
c) identifying rMVA virus expressing one or more of the IEfusion CMV protein antigens using one or more selection methods for serial passage;
d) conducting serial passage;
e) expanding an rMVA virus strain identified by step d); and
f) purifying the rMVA virus strain from step e) to form the composition.

5. The method of claim 4 , wherein the identification of rMVA virus carrying the transfer vector is accomplished by one or more gene-in selection methods, one or more gene-out selection methods, or a combination of gene-in and gene-out selection methods.

6. The method of claim 4 , wherein the serial passage is at least 10 passages.

7. The method of claim 4 , wherein the transfer plasmid vector further comprises the nucleotide sequence of SEQ ID NO:10.

8. The method of claim 4 , wherein the transfer plasmid comprises nucleotide sequences SEQ ID NO:9 and SEQ ID NO:10.

9. The method of claim 1 , wherein an immune response in the subject is modified.

10. The method of claim 9 , wherein the subject is a human.

11. The method of claim 9 , wherein the subject is a human stem cell donor or a human solid organ transplant donor.

12. The method of claim 9 , wherein the subject is a human with an immunodeficiency disease selected from the group consisting of HIV and a heritable immunodeficiency and the subject is susceptible to infection by human cytomegalovirus.

13. The method of claim 9 , wherein the subject is a human subject who has received a stem cell transplant or a solid organ transplant from a healthy donor.

Assignees

• Hope City

Inventors

• Diamond Don
• Wang Zhongde

Classifications

• A61K39/285 Primary

  Vaccinia virus or variola virus · CPC title

• C07K14/005

  from viruses · CPC title

• A61K39/245

  Herpetoviridae, e.g. herpes simplex virus · CPC title

• A61K39/12

  Viral antigens · CPC title

• C12N2710/24143

  viral genome or elements thereof as genetic vector · CPC title