Viral adjuvants
The present invention provides viral adjuvants for enhancing an immune response to an immunogen. In particular embodiments, the viral adjuvant is an alphavirus adjuvant or a Venezuelan Equine Encephalitis viral adjuvant. Also provided are compositions comprising the viral adjuvant and an immunogen, and pharmaceutical formulations comprising the viral adjuvant or compositions of the invention in a pharmaceutically acceptable carrier. Further provided are methods of producing an immune response against an immunogen in a subject comprising administering the immunogen and a viral adjuvant of the invention to the subject.
1. An alphavirus adjuvant comprising:
a modified alphavirus genomic nucleic acid that lacks sequences encoding the alphavirus structural proteins required for production of new alphavirus particles; wherein the modified alphavirus genome does not comprise a heterologous nucleic acid sequence that encodes a polypeptide of interest or a functional untranslated RNA and the 26S promoter is inactivated or deleted.
2. The alphavirus adjuvant of claim 1 , wherein the alphavirus adjuvant is replication-competent.
3. The alphavirus adjuvant of claim 1 , wherein the alphavirus adjuvant comprises a propagation-defective alphavirus particle that further comprises an alphavirus virion coat that packages the modified alphavirus genomic nucleic acid.
4. The alphavirus adjuvant of claim 1 , wherein the modified alphavirus genomic nucleic acid does not comprise a heterologous nucleic acid sequence.
5. The alphavirus adjuvant of claim 1 , wherein the alphavirus adjuvant is attenuated.
6. The alphavirus adjuvant of claim 1 , wherein the modified alphavirus genomic nucleic acid is a modified Venezuelan Equine Encephalitis (VEE) viral genomic nucleic acid.
7. The alphavirus adjuvant of claim 6 , wherein the alphavirus adjuvant comprises a propagation-defective VEE particle that further comprises a VEE virion coat that packages the VEE viral genomic nucleic acid.
8. The alphavirus adjuvant of claim 6 , wherein the modified alphavirus genomic nucleic acid does not comprise a heterologous nucleic acid sequence.
9. A pharmaceutical formulation comprising the alphavirus adjuvant of claim 1 in a pharmaceutically acceptable carrier.
10. A composition comprising the alphavirus adjuvant of claim 1 and an immunogen.
11. A pharmaceutical formulation comprising the composition of claim 10 in a pharmaceutically acceptable carrier.
12. A helper cell for producing an alphavirus adjuvant comprising an infectious propagation-defective alphavirus particle, comprising in an alphavirus-permissive cell:
(a) a modified alphavirus genomic RNA that comprises (i) sequences encoding the alphavirus nonstructural proteins, and (ii) a competent alphavirus packaging sequence, and (iii) wherein the modified alphavirus genomic nucleic acid lacks sequences encoding the alphavirus structural proteins required for production of new alphavirus particles; and further wherein the modified alphavirus genomic nucleic acid does not comprise a heterologous nucleic acid sequence that encodes a polypeptide of interest or a functional untranslated RNA and the 26S promoter is inactivated or deleted;
(b) helper nucleic acid(s) encoding the alphavirus structural proteins sufficient for producing an alphavirus particle;
wherein the combined expression of the modified alphavirus genomic RNA and the helper nucleic acid(s) encoding the alphavirus structural proteins produces an assembled alphavirus particle comprising the modified alphavirus genomic RNA.
13. The helper cell of claim 12 , wherein the helper nucleic acid(s) are RNA molecules that are transfected into the helper cell.
14. The helper cell of claim 12 , wherein the modified alphavirus genomic nucleic acid is a modified Venezuelan Equine Encephalitis (VEE) genomic nucleic acid.
15. The helper cell of claim 12 , wherein the alphavirus adjuvant comprises a VEE virion coat and the helper nucleic acid(s) encodes VEE structural proteins.
16. The helper cell of claim 12 , wherein the alphavirus adjuvant comprises infectious propagation-defective VEE particles, the modified alphavirus genomic nucleic acid is a modified VEE genomic nucleic acid, and the helper nucleic acid(s) encodes VEE structural proteins.
17. A method of making an alphavirus adjuvant comprising infectious propagation-defective alphavirus particles, comprising:
(a) providing a helper cell according to claim 12 ;
(b) producing the alphavirus particles in the helper cell; and
(c) collecting the alphavirus particles from the helper cell.
18. A pharmaceutical formulation comprising the alphavirus adjuvant of claim 6 in a pharmaceutically acceptable carrier.
19. A pharmaceutical formulation comprising the alphavirus adjuvant of claim 7 in a pharmaceutically acceptable carrier.
20. A composition comprising the alphavirus adjuvant of claim 6 and an immunogen.
21. A composition comprising the alphavirus adjuvant of claim 7 and an immunogen.