Compositions and vaccines comprising vesicles and methods of using the same
The disclosure relates to compositions, pharmaceutical compositions, vaccines, and methods of making and using the same. The compositions of the disclosure are useful to stimulate an antigen-specific immune response and, in some embodiments, a protective immune response in an animal after challenge to pathogens. The compositions of the disclosure are also useful for treatment and prevention of disease in a subject such as cancer.
1. A composition comprising a catanionic surfactant vesicle;
wherein the catanionic surfactant vesicle comprises a unilamellar bilayer comprising a mixture of oppositely charged single-tailed surfactants and one or a plurality of bacterial immunogens from whole bacterial cell extract;
wherein the at least one or plurality of bacterial immunogens comprise at least one membrane-bound sugar, or at least one membrane-bound polypeptide or at least one combination of a membrane-bound sugar and membrane-bound polypeptide; and
wherein the membrane-bound sugar, the membrane-bound polypeptide, or the combination of the membrane-bound sugar and membrane-bound polypeptide is positioned at least partially in the unilamellar bilayer; and
wherein the catanionic surfactant vesicle comprises bacterial lipooligosaccharide (LOS) or lipopolysaccharide (LPS) and one or a plurality of bacterial lipoproteins or bacterial membrane-bound protein; and
wherein the catanionic surfactant vesicle comprises a percentage of carbohydrate from about 0% to about 25% by weight.
2. The composition of claim 1 , wherein the at least one or plurality of bacterial immunogens is from a Gram-negative bacterial cell.
3. The composition of claim 1 , wherein the at least one or plurality of bacterial immunogens is derived from cells that are from Neisseria or Francisella.
4. The composition of claim 1 , wherein the catanionic surfactant vesicle comprises a percentage of carbohydrate from about 0.1% to about 16% by weight.
5. The composition of claim 1 , wherein the one or plurality of bacterial immunogens comprise one or a combination of pilin, porA, porB, OPA, DnaK, Tul4, and FopA.
6. The composition of claim 1 , wherein the one or plurality of bacterial immunogens comprises Pan HLA-DR reactive epitope (PADRE) individually or conjugated to one or a plurality of membrane-bound sugar molecules.
7. The composition of claim 1 , wherein the catanionic surfactant vesicle is lyophilized.
8. A pharmaceutical composition comprising: the composition of claim 1 and a pharmaceutically acceptable carrier.
9. A vaccine comprising:
(i) a plurality of catanionic vesicles, wherein at least one of the catanionic vesicles comprises a unilamellar bilayer comprising a mixture of oppositely charged single-tailed surfactants and one or a plurality of bacterial immunogens from whole bacterial cell extract wherein the one or plurality of bacterial immunogens comprise at least one membrane-bound sugar, or at least one membrane-bound polypeptide or at least one combination of a membrane-bound sugar and a polypeptide;
wherein the membrane-bound sugar, the membrane-bound polypeptide or the combination of the membrane-bound sugar and the membrane-bound polypeptide is positioned at least partially in the unilamellar bilayer, and
(ii) a pharmaceutically acceptable carrier;
wherein the one or plurality of bacterial immunogens is from Neisseria or Francisella cells; and
wherein the catanioinc vesicles comprise a diameter of 1000 nm or less.
10. The vaccine of claim 9 further comprising one or a plurality of: (i) a nucleic acid molecule encoding one or a plurality of immunogens; (ii) a nucleic acid molecule encoding one or a plurality of immunomodulating proteins; (iii) a protein adjuvant; (iv) a synthetic adjuvant; (v) an attenuated bacterial cell; (vi) a bacterial cell comprising a nucleic acid molecule encoding one or a plurality of immunogens; or (vii) a bacterial cell comprising a nucleic acid molecule encoding one or a plurality of immunomodulating proteins.
11. A method of treating and/or preventing establishment of a pathogen infection in a subject in need thereof, comprising: administering to the subject the pharmaceutical composition of claim 8 .
12. A method of immunizing a subject comprising:
administering to a subject in need thereof an amount of the vaccine of claim 9 effective to elicit an immune response.
13. The method of claim 12 , wherein the immune response is a protective, antigen-specific immune response.
14. A method of integrating a bacterial antigen into a vesicle comprising:
(a) centrifuging bacterial cells comprising at least one bacterial antigen;
(b) removing any bacterial growth medium;
(c) adding a weight/weight molar ratio of cationic surfactant and anionic surfactant of either about 30:70 or about 70:30 in aqueous solution to the bacterial cells;
(d) stirring resulting mixture at room temperature for at least about 30 minutes;
wherein the method does not comprise a step of purification or removal of bacterial sugar molecules prior to steps (a)-(d).
15. The method of claim 14 , wherein the cationic surfactant and anionic surfactant are sodium dodecyl benzene sulfonate (SDBS) and cetyltrimethylammonium tosylate (CTAT), respectively, at a molar weight ratio of about 70:30.
16. The method of claim 14 , wherein the bacterial cells are from Neisseria or Francisella ; or wherein the bacterial cells comprise recombinant vectors expressing one or a plurality of membrane-bound sugars and membrane-bound polypeptides or a combination of a membrane-bound sugar and a membrane-bound polypeptide from Neisseria or Francisella.
17. The method of claim 14 , wherein the aqueous solution comprises a marker or active agent.
18. The method of claim 14 , wherein the method further comprises filtering the vesicles through a size exclusion column or ion chromatography column.
19. A method of creating a library of catanionic vesicles comprising:
(a) adding a weight/weight molar ratio of cationic surfactant to anionic surfactant in aqueous solution of about 20:80 or about 80:20 to a population of bacterial cells in pelleted form;
(b) stirring resulting mixture at room temperature for at least about 30 minutes;
wherein the method does not comprise a step of purification prior to steps (a)-(b) or a step of removing lipopolysaccharides or lipooligosaccharides from the vesicles.
20. The method of claim 19 , wherein the cationic surfactant and anionic surfactant are SDBS and CTAT, respectively, at a molar weight ratio of about 70:30.
21. The method of claim 19 , wherein the bacterial cells are from Neisseria or Francisella ; or wherein the bacterial cells comprise recombinant vectors expressing one or a plurality of membrane-bound sugars and membrane polypeptides or a combination of a membrane-bound sugar and a membrane-bound polypeptide from Neisseria or Francisella.
22. The method of claim 19 , wherein the aqueous solution comprises one or more of a dye, a fluorescent protein, or a chemiluminescent agent.
23. The method of claim 14 , wherein the method further comprises filtering the vesicles through a size exclusion column or ion chromatography column.
24. A method of manufacturing an antibody against a bacterial antigen comprising injecting the composition of claim 1 into a subject and subsequently collecting its serum.
25. The vaccine of claim 9 , wherein the catanionic vesicles comprise a diameter of 300 nm or less and a carbohydrate percent from about 0.1% to about 15% by weight.
26. The method of claim 14 , wherein the bacterial cells are from Neisseria and Francisella.
27. The method of claim 19 , wherein the bacterial cells are from Neisseria or Francisella.
28. The method of claim 16 , wherein the membrane-bound polypeptide is from Neisseria.
29. The method of claim 16 , wherein the membrane-bound polypeptide is from Francisella.
30. The method of claim 21 , wherein the membrane-bound polypeptide is from Neisseria.
31. The method of claim 21 , wherein the membrane-bound polypeptide is from Francisella.