GNA1870-based vesicle vaccines for broad spectrum protection against diseases caused by
View Patent ↗The present invention generally provides methods and compositions for eliciting an immune response against Neisseria spp. bacteria in a subject, particularly against a Neisseria meningitidis serogroup B strain. check for moving around
1. A composition comprising:
isolated antigenic outer membrane vesicles (OMVs) or microvesicles (MVs) prepared from a first Neisseria meningitidis that is genetically modified:
(a) to comprise a mutation in a gene involved in biosynthesis or modification of lipid A of its lipopolysaccharide and
(b) to overexpress a meningococcal GNA1870 polypeptide, wherein the overexpression consists of the overexpression of a full-length meningococcal GNA1870 polypeptide in the OMVs or the MVs at a level that is greater than 3 to 10 times the level of the endogenous GNA1870 polypeptide expressed by unmodified parental Neisseria meningitidis from which the first genetically modified first Neisseria meningitidis is obtained,
wherein the OMVs or the MVs are prepared without using a detergent, and the composition when administered to a mammalian subject, elicits serum antibodies specific to the overexpressed GNA1870 polypeptide that are bactericidal against at least three Neisseria meningitidis strains that express a GNA1870 polypeptide and a heterologous meningococcal PorA protein, and
a pharmaceutically acceptable carrier.
2. The composition of claim 1 , wherein the lipopolysaccharide of the genetically modified Neisseria meningitidis has no detectable toxic activity of lipid A or decreased toxic activity of lipid A relative to its wild-type lipid A.
3. The composition of claim 1 , wherein the mutation comprises a mutation in htrB gene, msbB gene, or lpxK gene.
4. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is four or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
5. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is five or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
6. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is six or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
7. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is seven or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
8. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is eight or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
9. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is nine or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
10. The composition of claim 1 , wherein the overexpressed GNA1870 polypeptide is overexpressed at a level that is ten or more times greater than the level of the endogenous GNA1870 polypeptide expressed by the unmodified parental Neisseria meningitidis from which the first genetically modified Neisseria meningitidis is obtained.
11. The composition of claim 1 , wherein the composition further comprises:
isolated antigenic outer membrane vesicles (OMVs) or microvesicles (MVs) prepared from a second Neisseria meningitidis that is genetically modified to overexpress a meningococcal GNA1870 polypeptide, wherein the overexpression consists of the overexpression of a heterologous full-length meningococcal GNA1870 polypeptide in the OMVs or the MVs at a level that is greater than 3 times the level of the endogenous GNA1870 polypeptide expressed by unmodified parental Neisseria meningitidis from which the second genetically modified Neisseria meningitidis is obtained, wherein the OMVs or the MVs are prepared without using a detergent, and the composition when administered to a mammalian subject, elicits serum antibodies specific to the GNA1870 polypeptide overexpressed by the second genetically modified Neisseria meningitidis and wherein the antibodies are bactericidal against at least three Neisseria meningitidis strains that express a GNA1870 polypeptide and a heterologous meningococcal PorA protein, wherein the second Neisseria meningitidis bacterium is genetically diverse to the first Neisseria meningitidis bacterium.
12. The composition of claim 11 , wherein the first and the second Neisseria meningitidis are genetically diverse in that the two Neisseria meningitidis differ in at least one of serogroup, serotype, or subserotype.
13. The composition of claim 1 , wherein the first Neisseria meningitidis is genetically modified to produce at least two different meningococcal GNA1870 polypeptides of different variant groups.
14. The composition of claim 1 , wherein the first Neisseria meningitidis is genetically modified to disrupt the production of the endogenous GNA1870 polypeptide.
15. The composition of claim 11 , wherein the second Neisseria meningitidis is genetically modified to disrupt the production of the endogenous GNA1870 polypeptide.
16. A method of producing the composition of claim 1 , the method comprising:
culturing the genetically modified first Neisseria meningitidis;
preparing the OMVs or the MVs from the culture; and
combining the OMVs or the MVs with the pharmaceutically acceptable carrier to produce the composition.
17. A method of producing the composition of claim 2 , the method comprising:
culturing the genetically modified first Neisseria meningitidis;
preparing the OMVs or the MVs from the culture; and
combining the OMVs or the MVs with the pharmaceutically acceptable carrier to produce the composition.
18. A method of producing the composition of claim 10 , the method comprising:
culturing the genetically modified first Neisseria meningitidis;
preparing the OMVs or the MVs from the culture; and
combining the OMVs or the MVs with the pharmaceutically acceptable carrier to produce the composition.
19. A method of producing the composition of claim 11 , the method comprising:
culturing the genetically modified first and the second Neisseria meningitidis;
preparing the OMVs or the MVs from each of the cultures; and
combining the OMVs or the MVs with the pharmaceutically acceptable carrier to produce the composition.
20. A method of eliciting a bactericidal immune response against Neisseria meningitidis in a mammalian subject, the method comprising administering to the mammalian subject an immunologically effective amount of the composition of claim 1 .
21. A method of eliciting a bactericidal immune response against Neisseria meningitidis in a mammalian subject, the method comprising administering to the mammalian subject an immunologically effective amount of the composition of claim 2 .
22. A method of eliciting a bactericidal immune response against Neisseria meningitidis in a mammalian subject, the method comprising administering to the mammalian subject an immunologically effective amount of the composition of claim 4 .
23. A method of eliciting a bactericidal immune response against Neisseria meningitidis in a mammalian subject, the method comprising administering to the mammalian subject an immunologically effective amount of the composition of claim 10 .
24. A method of eliciting a bactericidal immune response against Neisseria meningitidis in a mammalian subject, the method comprising administering to the mammalian subject an immunologically effective amount of the composition of claim 11 .