Immunisation of large mammals with low doses of RNA
RNA encoding an immunogen is delivered to a large mammal at a dose of between 2 μg and 100 μg. Thus, the invention provides a method of raising an immune response in a large mammal, comprising administering to the mammal a dose of between 2 μg and 100 μg of immunogen-encoding RNA. Similarly, RNA encoding an immunogen can be delivered to a large mammal at a dose of 3 ng/kg to 150 ng/kg. The delivered RNA can elicit an immune response in the large mammal.
1. A method of eliciting an antibody response against an immunogen by an immune system in a large mammal, the method comprising administering to the large mammal at least two unit doses; each unit dose comprising a composition comprising liposomes and messenger ribonucleic acid (mRNA) molecules; the mRNA molecules comprising a poly-adenosine monophosphate (poly(A)) tail and a sequence that encodes the immunogen; the immunogen comprising a respiratory syncytial virus (RSV) surface fusion glycoprotein (F protein) immunogen, an Epstein-Barr virus (EBV) glycoprotein immunogen, a cytomegalovirus (CMV) glycoprotein immunogen, a human papillomavirus (HPV) 16 immunogen, a coronavirus spike polypeptide immunogen, an influenza virus immunogen, a Varicella zoster virus (VZV) glycoprotein immunogen, or a flavivirus immunogen; each unit dose comprising between 2 μg and 100 μg of the mRNA molecules; the at least two unit doses being sequential and at least 1 week apart; the administering comprising contacting the composition with skeletal muscle; the liposomes comprising a cationic lipid comprising a tertiary amine; the liposomes encapsulating at least half of the mRNA molecules; and the large mammal being a human, a horse, a cow, or a pig.
2. The method of claim 1 , the mRNA molecules further comprising a modified nucleotide.
3. The method of claim 2 , the liposomes further comprising: i) an anionic phospholipid or a zwitterionic phospholipid and ii) cholesterol.
4. The method of claim 3 , the method comprising administering to the large mammal at least three unit doses.
5. The method of claim 4 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
6. The method of claim 3 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
7. The method of claim 2 , the modified nucleotide including a modified pyrimidine nucleotide.
8. The method of claim 7 , the modified nucleotide further including a 5′ cap.
9. The method of claim 8 , the immunogen comprising the flavivirus immunogen; the liposomes further comprising DSPC and cholesterol; the large mammal being the human.
10. The method of claim 9 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
11. The method of claim 8 , the method comprising administering to the large mammal at least three unit doses.
12. The method of claim 11 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
13. The method of claim 8 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
14. The method of claim 7 , the immunogen comprising the flavivirus immunogen; the liposomes further comprising DSPC and cholesterol; the large mammal being the human.
15. The method of claim 14 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
16. The method of claim 7 , the method comprising administering to the large mammal at least three unit doses.
17. The method of claim 16 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
18. The method of claim 7 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
19. The method of claim 2 , the method comprising administering to the large mammal at least three unit doses.
20. The method of claim 19 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
21. The method of claim 2 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
22. The method of claim 1 , at least 80% of the liposomes having diameters from 20 nm to 220 nm.
23. The method of claim 22 , the method comprising administering to the large mammal at least three unit doses.
24. The method of claim 23 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
25. The method of claim 22 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
26. The method of claim 1 , the immunogen comprising the flavivirus immunogen; the liposomes further comprising 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol; the large mammal being the human.
27. The method of claim 26 , the method comprising administering to the large mammal at least three unit doses.
28. The method of claim 27 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
29. The method of claim 26 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
30. The method of claim 1 , the method comprising administering to the large mammal at least three unit doses.
31. The method of claim 30 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
32. The method of claim 1 , the mRNA molecules being self-replicating RNA molecules.
33. The method of claim 32 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
34. The method of claim 1 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
35. A method of eliciting an antibody response against an immunogen by an immune system in a large mammal, the method comprising administering to the large mammal at least two unit doses, each unit dose comprising a composition comprising liposomes and mRNA molecules; the mRNA molecules comprising a poly(A) tail and a sequence that encodes the immunogen; the immunogen comprising a RSV F protein immunogen, an EBV glycoprotein immunogen, a CMV glycoprotein immunogen, a HPV16 immunogen, a coronavirus spike polypeptide immunogen, an influenza virus immunogen, a VZV glycoprotein immunogen, or a flavivirus immunogen; each unit dose comprising between 0.5 μg and 1.5 μg of the mRNA molecules per kg of the body mass of the large mammal; the at least two unit doses being sequential and at least 1 week apart; the administering comprising contacting the composition with skeletal muscle; the liposomes comprising a cationic lipid comprising a tertiary amine; the liposomes encapsulating at least half of the mRNA molecules; and the large mammal being a human, a horse, a cow, or a pig.
36. The method of claim 35 , the mRNA molecules further comprising a modified nucleotide.
37. The method of claim 36 , the liposomes further comprising: i) an anionic phospholipid or a zwitterionic phospholipid and ii) cholesterol.
38. The method of claim 37 , the method comprising administering to the large mammal at least three unit doses.
39. The method of claim 38 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
40. The method of claim 37 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
41. The method of claim 36 , the modified nucleotide including a modified pyrimidine nucleotide.
42. The method of claim 41 , the modified nucleotide further including a 5′ cap.
43. The method of claim 42 , the immunogen comprising the flavivirus immunogen; the liposomes further comprising DSPC and cholesterol; the large mammal being the human.
44. The method of claim 43 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
45. The method of claim 42 , the method comprising administering to the large mammal at least three unit doses.
46. The method of claim 45 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
47. The method of claim 42 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
48. The method of claim 41 , the immunogen comprising the flavivirus immunogen; the liposomes further comprising DSPC and cholesterol; the large mammal being the human.
49. The method of claim 48 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
50. The method of claim 41 , the method comprising administering to the large mammal at least three unit doses.
51. The method of claim 50 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
52. The method of claim 41 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
53. The method of claim 36 , the method comprising administering to the large mammal at least three unit doses.
54. The method of claim 53 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
55. The method of claim 36 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
56. The method of claim 35 , at least 80% of the liposomes having diameters from 20 nm to 220 nm.
57. The method of claim 56 , the method comprising administering to the large mammal at least three unit doses.
58. The method of claim 57 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
59. The method of claim 56 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
60. The method of claim 35 , the immunogen comprising the flavivirus immunogen; the liposomes further comprising cholesterol and DSPC; the large mammal being the human.
61. The method of claim 60 , the method comprising administering to the large mammal at least three unit doses.
62. The method of claim 61 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
63. The method of claim 60 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
64. The method of claim 35 , the method comprising administering to the large mammal at least three unit doses.
65. The method of claim 64 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
66. The method of claim 35 , the mRNA molecules being self-replicating RNA molecules.
67. The method of claim 66 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.
68. The method of claim 35 , the method further eliciting a cell-mediated immune response against the immunogen by the immune system in the large mammal.