Diatom-based vaccines

Patent metadata

Field	Value
Publication number	US-9358283-B2
Application number	US-201214353721-A
Country	US
Kind code	B2
Filing date	Oct 26, 2012
Priority date	Oct 28, 2011
Publication date	Jun 7, 2016
Grant date	Jun 7, 2016

Abstract

This invention provides diatom-based vaccines.

First claim

What is claimed is:

1. A diatom comprising a nucleic acid encoding an antigen heterologous to the diatom, wherein the antigen is expressed as a fusion protein with a surface-expressed polypeptide endogenous to the diatom selected from a frustulin and p150 cell surface protein, wherein the fusion protein is attached to the surface of the diatom, wherein the diatom is an intact cell.

2. The diatom of claim 1 , wherein the antigen is expressed as a fusion protein with a frustulin protein having at least 90% sequence identity to SEQ ID NO:6 or SEQ ID NO:7.

3. The diatom of claim 1 , wherein the antigen is expressed under the control of a promoter endogenous to the diatom selected from the group consisting of fucoxanthin chlorophyll binding protein (FCP) promoter and ribosomal protein L41 (rpL41) promoter.

4. The diatom of claim 3 , wherein the FCP promoter comprises a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:1.

5. The diatom of claim 1 , wherein the antigen is an antibody or fragment thereof.

6. The diatom of claim 5 , wherein the antibody is a transmission blocking antibody, wherein the transmission blocking antibody inhibits the growth and/or replication of an infectious agent.

7. The diatom of claim 5 , wherein the antibody is a single domain antibody or a nanobody.

8. The diatom of claim 1 , wherein the diatom is a Thalassiosira pseudonana.

9. An expression cassette comprising:
i) a promoter selected from the group consisting of fucoxanthin chlorophyll binding protein (FCP) promoter and ribosomal protein L41 (rpL41) promoter; the promoter operably linked to;
ii) a nucleic acid encoding a diatom cell surface polypeptide selected from a frustulin and p150 cell surface protein; the nucleic acid encoding the diatom cell surface polypeptide operably linked to;
iii) a nucleic acid encoding an antigen heterologous to a diatom.

10. The expression cassette of claim 9 , wherein the FCP promoter comprises a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:1.

11. The expression cassette of claim 9 , wherein the antigen is expressed as a fusion protein with a frustulin protein having at least 90% sequence identity to SEQ ID NO:6 or SEQ ID NO:7.

12. The expression cassette of claim 9 , wherein the antigen is an antibody or fragment thereof.

13. The expression cassette of claim 2 , wherein the antibody is a transmission blocking antibody, wherein the transmission blocking antibody inhibits the growth and/or replication of an infectious agent.

14. The expression cassette of claim 12 , wherein the antibody is a single domain antibody or a nanobody.

15. A vector or plasmid comprising the expression cassette of claim 9 .

16. The vector or plasmid of claim 15 , wherein the vector is a plasmid comprising a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:8.

17. A diatom comprising the expression cassette of claim 9 .

18. The diatom of claim 17 , wherein the diatom is a Thalassiosira pseudonana.

19. A composition comprising a diatom or a population of diatoms of claim 1 and a pharmaceutically acceptable carrier.

20. A method of stimulating an immune response in an animal, comprising administering to the animal a composition of claim 19 in an amount effective to stimulate the immune response in the animal.

21. The method of claim 20 , wherein the diatom is administered in the water or food or by aerosol.

22. The method of claim 20 , wherein the animal is a mammal.

23. A sonicated diatom particle attached to a fusion protein, the fusion protein comprises an antigen heterologous to the diatom fused to a surface-expressed polypeptide endogenous to the diatom selected from a frustulin and p150 cell surface protein.

24. The sonicated diatom particle of claim 23 , wherein the frustulin protein has at least 90% sequence identity to SEQ ID NO:6 or SEQ ID NO:7.

25. The sonicated diatom particle of claim 23 , wherein the antigen is an antibody or fragment thereof.

26. The sonicated diatom particle of claim 25 , wherein the antibody is a transmission blocking antibody, wherein the transmission blocking antibody inhibits the growth and/or replication of an infectious agent.

27. The sonicated diatom particle of claim 25 , wherein the antibody is a single domain antibody or a nanobody.

28. The sonicated diatom particle of claim 23 , wherein the diatom particle is from a Thalassiosira pseudonana.

29. The sonicated diatom particle of claim 23 , wherein the diatom particle is a microparticle or a nanoparticle.

30. A composition comprising a sonicated diatom or a population of sonicated diatom particles of claim 23 and a pharmaceutically acceptable carrier.

31. A method of stimulating an immune response in an animal, comprising administering to the animal a composition of claim 30 in an amount effective to stimulate the immune response in the animal.

32. The method of claim 31 , wherein the sonicated diatom particle is administered in the water or food or by aerosol.

33. The method of claim 31 , wherein the animal is a mammal.

Assignees

• Univ California
• Synaptic Res Llc

Inventors

• Corbeil Lynette B
• Hildebrand Mark
• Shrestha Roshan
• Davis Aubrey
• Schrier Rachel
• Oyler George A
• Rosenberg Julian N

Classifications

• C12N1/12 Primary

  Unicellular algae; Culture media therefor (as new plants A01H13/00) · CPC title

• A61K2039/523

  expressing foreign proteins · CPC title

• A61K39/102

  {Pasteurellales, e.g. Actinobacillus}, Pasteurella; Haemophilus · CPC title

• A61K2039/542

  oral/gastrointestinal · CPC title

• C12N15/79

  Vectors or expression systems specially adapted for eukaryotic hosts · CPC title