Salmonella typhi Ty21a expressing Yersinia pestis F1-V fusion protein and uses thereof

Patent metadata

Field	Value
Publication number	US-9409956-B2
Application number	US-201314402457-A
Country	US
Kind code	B2
Filing date	May 22, 2013
Priority date	May 23, 2012
Publication date	Aug 9, 2016
Grant date	Aug 9, 2016

Abstract

Described herein is the generation of a plasmid construct for expression of a Yersinia pestis F1-V fusion protein. In the disclosed plasmid, the F1-V fusion protein coding sequence is operably linked to the E. coli htrA promoter, and is fused in-frame to the E. coli HlyA secretion signal sequence. Also described is Salmonella enterica serovar Typhi strain Ty21a containing the F1-V fusion protein expression plasmid, such as for use as an oral vaccine against plague.

First claim

The invention claimed is:

1. A recombinant nucleic acid molecule comprising a Y. pestis F1-V fusion protein coding sequence operably linked to an E. coli htrA promoter sequence, and further comprising an E. coli HlyA secretion signal peptide secretion signal peptide coding sequence fused in-frame with the F1-V fusion protein coding sequence, wherein the sequence of the nucleic acid molecule comprises SEQ ID NO: 1.

2. A plasmid comprising the recombinant nucleic acid molecule of claim 1 , an origin of replication, a selectable marker and the E. coli HlyB and HlyD protein coding sequences.

3. A plasmid comprising the recombinant nucleic acid molecule of claim 1 .

4. A recombinant nucleic acid molecule comprising a Yersinia pestis F1-V fusion protein coding sequence operably linked to an Escherichia coli htrA promoter sequence, wherein the nucleic acid molecule comprises a sequence at least 95% identical to nucleotides 1-1558 of SEQ ID NO: 1.

5. The recombinant nucleic acid molecule of claim 4 , wherein the nucleic acid molecule comprises the sequence of nucleotides 1-1558 of SEQ ID NO: 1.

6. The recombinant nucleic acid molecule of claim 4 , further comprising a secretion signal peptide coding sequence fused in-frame with the F1-V fusion protein coding sequence.

7. The recombinant nucleic acid molecule of claim 6 , wherein the secretion signal peptide is the E. coli HlyA secretion signal peptide.

8. The recombinant nucleic acid molecule of claim 6 , wherein the sequence of the nucleic acid molecule is at least 95% identical to SEQ ID NO: 1.

9. A plasmid comprising the recombinant nucleic acid molecule of claim 4 .

10. The plasmid of claim 9 , wherein the plasmid is a low-copy number plasmid.

11. The plasmid of claim 9 , further comprising a selectable marker.

12. The plasmid of claim 11 , wherein the selectable marker is an antibiotic resistance gene.

13. The plasmid of claim 9 , further comprising the E. coli HlyB and HlyD protein coding sequences.

14. A Salmonella Typhi Ty21a bacterium comprising the recombinant nucleic acid molecule of claim 4 .

15. An immunogenic composition comprising the Salmonella Typhi Ty21a bacterium of claim 14 and a pharmaceutically acceptable carrier.

16. The immunogenic composition of claim 15 , wherein the composition is formulated for oral administration.

17. A method of eliciting an immune response against Yersinia pestis in a subject, comprising administering to the subject the Salmonella Typhi Ty21a bacterium of claim 14 .

18. The method of claim 17 , wherein the immune response is a protective immune response.

19. A method of reducing the risk of developing plague or treating plague in a subject, comprising administering to the subject the Salmonella Typhi Ty21a of claim 14 .

Assignees

• Us Health

Inventors

• Kopecko Dennis J
• Osorio Manuel
• Foote Monica R

Classifications

• A61K39/0291

  Yersinia · CPC title

• C07K2319/40

  containing a tag for immunodetection, or an epitope for immunisation · CPC title

• C07K2319/036

  targeting to the medium outside of the cell, e.g. type III secretion · CPC title

• A61K2039/523

  expressing foreign proteins · CPC title

• C07K14/24 Primary

  from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia · CPC title