Delivery of RNA to trigger multiple immune pathways
RNA encoding an immunogen is co-delivered to non-immune cells at the site of delivery and also to immune cells which infiltrate the site of delivery. The responses of these two cell types to the same delivered RNA lead to two different effects, which interact to produce a strong immune response against the immunogen. The non-immune cells translate the RNA and express the immunogen. Infiltrating immune cells respond to the RNA by expressing type I interferons and pro-inflammatory cytokines which produce a local adjuvant effect which acts on the immunogen-expressing non-immune cells to upregulate major histocompatibility complex expression, thereby increasing presentation of the translated protein to T cells. The effects on the immune and non-immune cells can be achieved by a single delivery of a single RNA e.g. by a single injection.
1. An immunogenic composition comprising lipid particles and messenger ribonucleic acid (mRNA) molecules; the mRNA molecules comprising a 7′-methylguanosine, a first 5′ ribonucleotide, and a sequence that encodes an immunogen; the immunogen comprising a respiratory syncytial virus (RSV) immunogen, an Epstein-Barr virus (EBV) immunogen, a cytomegalovirus (CMV) immunogen, a coronavirus spike polypeptide immunogen, an influenza virus immunogen, a Varicella zoster virus (VZV) immunogen, or a flavivirus immunogen; the first 5′ ribonucleotide comprising a 2′-methylated ribose; the 7′ methylguanosine linked 5′-to-5′ to the first 5′ ribonucleotide; the lipid particles comprising a polyethylene glycol-ylated (PEGylated) lipid, cholesterol, a first phospholipid, and a cationic lipid; the cationic lipid comprising a tertiary amine; the first phospholipid comprising an anionic phospholipid or a zwitterionic phospholipid; the lipid particles encapsulating at least half of the mRNA molecules; and the immunogenic composition being immunogenic in vivo by eliciting at least: (i) an antibody response against the immunogen, (ii) a cell-mediated immune response against the immunogen, or (iii) both (i) and (ii).
2. The immunogenic composition of claim 1 , the mRNA molecules comprising modified nucleotides.
3. The immunogenic composition of claim 2 , the mRNA molecules further comprising a poly(adenosine monophosphate) (poly(A)) tail.
4. The immunogenic composition of claim 3 , the mRNA molecules being positive-stranded.
5. The immunogenic composition of claim 1 , the mRNA molecules being a self-replicating RNA.
6. An immunogenic composition comprising lipid particles and mRNA molecules; the mRNA molecules comprising a 7′-methylguanosine, a first 5′ ribonucleotide, and a sequence that encodes an immunogen; the first 5′ ribonucleotide comprising a 2′-methylated ribose; the 7′ methylguanosine linked 5′-to-5′ to the first 5′ ribonucleotide; the lipid particles comprising a PEGylated lipid, cholesterol, a first phospholipid, and a cationic lipid; the cationic lipid comprising a tertiary amine; the first phospholipid comprising an anionic phospholipid or a zwitterionic phospholipid; the lipid particles encapsulating at least half of the mRNA molecules; and the immunogenic composition being immunogenic in vivo by eliciting at least: (i) an antibody response against the immunogen, (ii) a cell-mediated immune response against the immunogen, or (iii) both (i) and (ii).
7. The immunogenic composition of claim 6 , the mRNA molecules comprising modified nucleotides.
8. The immunogenic composition of claim 7 , the mRNA molecules further comprising a poly(A) tail.
9. The immunogenic composition of claim 8 , the mRNA molecules being positive-stranded.
10. The immunogenic composition of claim 6 , the mRNA molecules being a self-replicating RNA.
11. A composition comprising liposomes and mRNA molecules; the mRNA molecules comprising a 7′-methylguanosine, a first 5′ ribonucleotide, a poly(A) tail, and a sequence that encodes an immunogen; the immunogen comprising a RSV immunogen, an EBV immunogen, a CMV immunogen, a coronavirus spike polypeptide immunogen, an influenza virus immunogen, a VZV immunogen, or a flavivirus immunogen; the first 5′ ribonucleotide comprising a 2′-methylated ribose; the 7′ methylguanosine linked 5′-to-5′ to the first 5′ ribonucleotide; the liposomes comprising a cationic lipid; the cationic lipid comprising a tertiary amine; and the liposomes encapsulating at least half of the mRNA molecules.
12. The composition of claim 11 , the mRNA molecules comprising modified nucleotides.
13. The composition of claim 12 , the mRNA molecules being positive-stranded.
14. The composition of claim 11 , the mRNA molecules being a self-replicating RNA.
15. The composition of claim 13 , the liposomes further comprising cholesterol and a first phospholipid comprising an anionic phospholipid or a zwitterionic phospholipid.
16. A composition comprising liposomes and mRNA molecules; the mRNA molecules comprising a 7′-methylguanosine, a first 5′ ribonucleotide, poly(A) tail, and a sequence that encodes an immunogen; the immunogen comprising a bacterial immunogen, a viral immunogen, a fungal immunogen, or a parasite immunogen; the first 5′ ribonucleotide comprising a 2′-methylated ribose; the 7′ methylguanosine linked 5′-to-5′ to the first 5′ ribonucleotide; the liposomes comprising a cationic lipid; the cationic lipid comprising a tertiary amine; and the liposomes encapsulating at least half of the mRNA molecules.
17. The composition of claim 16 , the mRNA molecules comprising modified nucleotides.
18. The composition of claim 16 , the mRNA molecules being positive-stranded.
19. The composition of claim 16 , the mRNA molecules being a self-replicating RNA.
20. The composition of claim 16 , the liposomes further comprising cholesterol and a first phospholipid comprising an anionic phospholipid or a zwitterionic phospholipid.
21. A method of eliciting an immune response in a human, the method comprising administering to the human an effective amount to elicit the immune response of the immunogenic composition of claim 4 ; the immune response comprising the antibody response against the immunogen or the cell-mediated immune response against the immunogen.
22. The method of claim 21 comprising administering at least two unit doses of the immunogenic composition to the human.
23. The method of claim 21 comprising administering the immunogenic composition to skeletal muscle tissue by injection.
24. A method of eliciting an immune response in a vertebrate, the method comprising administering to the vertebrate an effective amount to elicit the immune response of the immunogenic composition of claim 6 ; the immune response comprising the antibody response against the immunogen or the cell-mediated immune response against the immunogen.
25. The method of claim 24 comprising administering at least two unit doses of the immunogenic composition to the vertebrate.
26. The method of claim 24 comprising administering the immunogenic composition to skeletal muscle tissue by injection.
27. A method of eliciting an immune response comprising an antibody response or a cell-mediated immune response in a human, the method comprising administering to the human an effective amount to elicit the immune response of the composition of claim 13 .
28. The method of claim 27 comprising administering at least two unit doses of the composition to the human.
29. A method of eliciting an immune response comprising an antibody response or a cell-mediated immune response in a vertebrate, the method comprising administering to the vertebrate an effective amount to elicit the immune response of the composition of claim 16 .
30. The method of claim 29 comprising administering at least two unit doses of the composition to the vertebrate.
31. The method of claim 29 , the vertebrate being a human.
32. The method of claim 24 , the vertebrate being a human.
33. The immunogenic composition of claim 4 , the immunogen comprising the RSV immunogen.
34. The immunogenic composition of claim 4 , the immunogen comprising the EBV immunogen.
35. The immunogenic composition of claim 4 , the immunogen comprising the CMV immunogen.
36. The immunogenic composition of claim 4 , the immunogen comprising the coronavirus spike polypeptide immunogen.
37. The immunogenic composition of claim 4 , the immunogen comprising the influenza virus immunogen.
38. The immunogenic composition of claim 37 , the influenza virus immunogen comprising an influenza virus A immunogen.
39. The immunogenic composition of claim 4 , the immunogen comprising the VZV immunogen.
40. The immunogenic composition of claim 4 , the immunogen comprising the flavivirus immunogen.
41. The immunogenic composition of claim 33 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).
42. The immunogenic composition of claim 34 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
43. The immunogenic composition of claim 35 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
44. The immunogenic composition of claim 36 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
45. The immunogenic composition of claim 37 the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
46. The immunogenic composition of claim 38 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
47. The immunogenic composition of claim 39 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
48. The immunogenic composition of claim 40 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
49. The immunogenic composition of claim 9 , the first phospholipid comprising a zwitterionic lipid; the zwitterionic lipid comprising DSPC.
50. The composition of claim 13 , the immunogen comprising the RSV immunogen.
51. The composition of claim 13 , the immunogen comprising the EBV immunogen.
52. The composition of claim 13 , the immunogen comprising the CMV immunogen.
53. The composition of claim 13 , the immunogen comprising the coronavirus spike polypeptide immunogen.
54. The composition of claim 13 , the immunogen comprising the influenza virus immunogen.
55. The composition of claim 54 , the influenza virus immunogen comprising an influenza virus A immunogen.
56. The composition of claim 13 , the immunogen comprising the VZV immunogen.
57. The composition of claim 13 , the immunogen comprising the flavivirus immunogen.
58. The composition of claim 50 , the liposomes further comprising cholesterol and DSPC.
59. The composition of claim 51 , the liposomes further comprising cholesterol and DSPC.
60. The composition of claim 52 , the liposomes further comprising cholesterol and DSPC.
61. The composition of claim 53 , the liposomes further comprising cholesterol and DSPC.
62. The composition of claim 54 , the liposomes further comprising cholesterol and DSPC.
63. The composition of claim 55 , the liposomes further comprising cholesterol and DSPC.
64. The composition of claim 56 , the liposomes further comprising cholesterol and DSPC.
65. The composition of claim 57 , the liposomes further comprising cholesterol and DSPC.
66. The composition of claim 18 , the liposomes further comprising cholesterol and DSPC.
67. The method of claim 21 , the immunogen comprising the RSV immunogen.
68. The method of claim 21 , the immunogen comprising the EBV immunogen.
69. The method of claim 21 , the immunogen comprising the CMV immunogen.
70. The method of claim 21 , the immunogen comprising the coronavirus spike polypeptide immunogen.
71. The method of claim 21 , the immunogen comprising the influenza virus immunogen.
72. The method of claim 71 , the influenza virus immunogen comprising an influenza virus A immunogen.
73. The method of claim 21 , the immunogen comprising the flavivirus immunogen.
74. The method of claim 21 , the immunogen comprising the VZV immunogen.
75. The method of claim 67 comprising administering at least two unit doses of the immunogenic composition to the human.
76. The method of claim 68 comprising administering at least two unit doses of the immunogenic composition to the human.
77. The method of claim 69 comprising administering at least two unit doses of the immunogenic composition to the human.
78. The method of claim 70 comprising administering at least two unit doses of the immunogenic composition to the human.
79. The method of claim 71 comprising administering at least two unit doses of the immunogenic composition to the human.
80. The method of claim 72 comprising administering at least two unit doses of the immunogenic composition to the human.
81. The method of claim 73 comprising administering at least two unit doses of the immunogenic composition to the human.
82. The method of claim 74 comprising administering at least two unit doses of the immunogenic composition to the human.
83. The method of claim 75 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
84. The method of claim 76 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
85. The method of claim 77 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
86. The method of claim 78 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
87. The method of claim 79 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
88. The method of claim 80 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
89. The method of claim 81 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
90. The method of claim 82 , the first phospholipid comprising the zwitterionic phospholipid; the zwitterionic phospholipid comprising DSPC.
91. The method of claim 27 , the immunogen comprising the RSV immunogen.
92. The method of claim 27 , the immunogen comprising the EBV immunogen.
93. The method of claim 27 , the immunogen comprising the CMV immunogen.
94. The method of claim 27 , the immunogen comprising the coronavirus spike polypeptide immunogen.
95. The method of claim 27 , the immunogen comprising the influenza virus immunogen.
96. The method of claim 91 , the influenza virus immunogen comprising an influenza virus A immunogen.
97. The method of claim 27 , the immunogen comprising the flavivirus immunogen.
98. The method of claim 27 , the immunogen comprising the VZV immunogen.
99. The method of claim 91 comprising administering at least two unit doses of the composition to the human.
100. The method of claim 92 comprising administering at least two unit doses of the composition to the human.
101. The method of claim 93 comprising administering at least two unit doses of the composition to the human.
102. The method of claim 94 comprising administering at least two unit doses of the composition to the human.
103. The method of claim 95 comprising administering at least two unit doses of the composition to the human.
104. The method of claim 96 comprising administering at least two unit doses of the composition to the human.
105. The method of claim 97 comprising administering at least two unit doses of the composition to the human.
106. The method of claim 98 comprising administering at least two unit doses of the composition to the human.
107. The method of claim 99 , the liposomes comprising cholesterol and DSPC.
108. The method of claim 100 , the liposomes comprising cholesterol and DSPC.
109. The method of claim 101 , the liposomes comprising cholesterol and DSPC.
110. The method of claim 102 , the liposomes comprising cholesterol and DSPC.
111. The method of claim 103 , the liposomes comprising cholesterol and DSPC.
112. The method of claim 104 , the liposomes comprising cholesterol and DSPC.
113. The method of claim 105 , the liposomes comprising cholesterol and DSPC.
114. The method of claim 106 , the liposomes comprising cholesterol and DSPC.