Generation of virus-like particles and use as panfilovirus vaccine
View Patent ↗In this application are described filovirus-like particles for both Ebola and Marburg and their use as a diagnostic and therapeutic agent as well as a filovirus vaccine. Also described is the association of Ebola and Marburg with lipid rafts during assembly and budding, and the requirement of functional rafts for entry of filoviruses into cells.
1. A filovirus virus like particle (VLP) comprising filovirus envelope glycoprotein (GP) and filovirus matrix protein VP40.
2. A filovirus VLP, produced by expressing in a cell a polynucleotide encoding filovirus envelope glycoprotein and filovirus matrix protein VP40 such that said polynucleotide is expressed and said VLP is produced.
3. A VLP of claim 1 where said filovirus is chosen from the group consisting of Ebola and Marburg.
4. A VLP of claim 2 where said filovirus is chosen from the group consisting of Ebola and Marburg.
5. A filovirus vaccine comprising VLP according to claim 1 .
6. A filovirus vaccine comprising VLP according to claim 2 .
7. A filovirus vaccine according to claim 6 further comprising an adjuvant.
8. The vaccine of claim 7 wherein said adjuvant is chosen from the group consisting of: RIBI, QS21 and LT(R192G).
9. A filovirus vaccine according to claim 5 wherein said filovirus is chosen from the group consisting of Ebola and Marburg.
10. A filovirus vaccine according to claim 6 wherein said filovirus is chosen from the group consisting of Ebola and Marburg.
11. A filovirus vaccine comprising VLP according to claim 1 and a nucleic acid encoding an agent capable of eliciting an immune response against said filovirus.
12. An Ebola VLP-producing cell comprising a mammalian cell expressing Ebola GP and VP40.
13. A kit for the detection of Ebola virus infection comprising Ebola VLPs according to claim 3 .
14. A kit for the detection of Marburg virus infection comprising Marburg VLPs according to claim 3 .
15. A kit for testing agents involved in Ebola budding said kit comprising a cell producing Ebola VLPs according to claim 12 and ancillary reagents for detecting VLPs in the supernatant of said cells when cells are cultured.
16. A Marburg VLP-producing cell comprising a mammalian cell expressing Marburg GP and VP40.
17. A kit for testing agents involved in Marburg budding said kit comprising a cell producing Marburg VLPs according to claim 16 and ancillary reagents for detecting VLPs in the supernatant of said cells when cells are cultured.
18. An immunogenic composition comprising, in a physiologically acceptable vehicle, Ebola VLPs according to claim 4 .
19. The immunogenic composition according to claim 18 which further comprises an adjuvant to enhance the immune response.
20. The immunogenic composition of claim 18 , wherein said Ebola VLPs are produced by expressing in a mammalian cell Ebola GP and Ebola VP40.
21. An immunogenic composition comprising, in a physiologically acceptable vehicle, Marburg VLPs according to claim 4 .
22. The immunogenic composition according to claim 21 which induces a Marburg specific immune response in a subject.
23. The immunogenic composition according to claim 21 , which further comprises an adjuvant to enhance the immune response.
24. The immunogenic composition of claim 21 , wherein said Marburg VLPs are produced by expressing in a mammalian cell Marburg GP and Marburg VP40.
25. A panfilovirus vaccine comprising a mixture of EBOV and MARV VLPs according to claim 4 .
26. A MARV vaccine protective against infection with MARV-Musoke, MARV-Ravn, and MARV-Ci67, comprising MARV VLPs according to claim 4 consisting essentially of GP and VP40 from MARV-Musoke.
27. An Ebola VLP producing cell comprising an insect cell expressing Ebola GP and VP40.
28. A Marburg VLP producing cell comprising an insect cell expressing Marburg GP and VP40.
29. A filovirus virus like particle (VLP) comprising filovirus envelope glycoprotein (GP), filovirus matrix protein VP40, and filovirus nucleoprotein (NP).
30. A filovirus VLP, produced by expressing in a cell a polynucleotide encoding filovirus envelope glycoprotein, filovirus matrix protein VP40, and filovirus nucleoprotein NP, such that said polynucleotide is expressed and said VLP is produced.
31. The filovirus VLP according to claim 30 wherein said cell is chosen from the group consisting of mammalian cell and insect cell.
32. A VLP of claim 29 where said filovirus is chosen from the group consisting of Ebola and Marburg.
33. A VLP of claim 30 where said filovirus is chosen from the group consisting of Ebola and Marburg.
34. A filovirus vaccine comprising VLP according to claim 29 .
35. A filovirus vaccine comprising VLP according to claim 30 .
36. A filovirus vaccine according to claim 34 further comprising an adjuvant.
37. The vaccine of claim 36 wherein said adjuvant is chosen from the group consisting of: RIBI, QS21 and LT(R192G).
38. A filovirus vaccine according to claim 34 wherein said filovirus is chosen from the group consisting of Ebola and Marburg.
39. A filovirus vaccine according to claim 35 wherein said filovirus is chosen from the group consisting of Ebola and Marburg.
40. A filovirus vaccine comprising VLP according to claim 29 and a nucleic acid encoding an agent capable of eliciting an immune response against said filovirus.
41. An Ebola VLP-producing cell wherein said cell expresses Ebola GP, VP40, and NP.
42. A method for detecting Ebola virus infection comprising contacting a sample from a subject suspected of having Ebola virus infection with an Ebola VLP according to claim 32 and detecting the presence or absence of an infection by detecting the presence or absence of a complex formed between the Ebola VLP and antibodies specific therefor in said sample.
43. A kit for the detection of Ebola virus infection comprising Ebola VLPs according to claim 32 .
44. A method for detecting Marburg virus infection comprising contacting a sample from a subject suspected of having Marburg virus infection with a Marburg VLP according to claim 32 and detecting the presence or absence of an infection by detecting the presence or absence of a complex formed between the Marburg VLP and antibodies specific therefor in said sample.
45. A kit for the detection of Marburg virus infection comprising Marburg VLPs according to claim 32 .
46. A kit for testing agents involved in Ebola budding said kit comprising a cell producing Ebola VLPs according to claim 41 and ancillary reagents for detecting VLPs in the supernatant of said cells when cells are cultured.
47. A Marburg VLP-producing cell wherein said cell expresses Marburg GP, VP40, and NP.
48. A kit for testing agents involved in Marburg budding said kit comprising a cell producing Marburg VLPs according to claim 47 and ancillary reagents for detecting VLPs in the supernatant of said cells when cells are cultured.
49. An immunogenic composition comprising, in a physiologically acceptable vehicle, Ebola VLPs according to claim 32 .
50. The immunogenic composition according to claim 49 which further comprises an adjuvant to enhance the immune response.
51. The immunogenic composition of claim 50 , wherein said Ebola VLPs are produced by expressing in an insect cell Ebola GP, Ebola VP40, and Ebola NP.
52. An immunogenic composition comprising, in a physiologically acceptable vehicle, Marburg VLPs according to claim 32 .
53. The immunogenic composition according to claim 52 which induces a Marburg specific immune response in a subject.
54. The immunogenic composition according to claim 52 , which further comprises an adjuvant to enhance the immune response.
55. The immunogenic composition of claim 52 , wherein said Marburg VLPs are produced by expressing in an insect cell Marburg GP, Marburg VP40, and Marburg NP.
56. A panfilovirus vaccine comprising a mixture of EBOV and MARV VLPs according to claim 32 .
57. The immunogenic composition according to claim 18 which induces an Ebola specific immune response in a subject.
58. The filovirus VLP according to claim 31 wherein said filovirus is chosen from the group consisting of Ebola and Marburg.