Delivery of RNA to trigger multiple immune pathways
RNA encoding an immunogen is co-delivered to non-immune cells as 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. A composition comprising lipid particles and messenger ribonucleic acid (mRNA) molecules; the mRNA molecules comprising: (i) a 5′ cap nucleoside, (ii) a first 5′ ribonucleoside, (iii) a triphosphate bridge, and (iv) a sequence that encodes a respiratory syncytial virus (RSV) surface fusion glycoprotein (F-protein) immunogen; the first 5′ ribonucleoside comprising a 2′-methylated ribose; the 5′ cap nucleoside linked 5′-to-5′ to the first 5′ ribonucleoside by the triphosphate bridge; the lipid particles comprising: (a) a polyethylene glycol-ylated lipid, (b) cholesterol, (c) an anionic phospholipid or a zwitterionic phospholipid, and (d) a cationic lipid comprising a tertiary amine; and the lipid particles encapsulating at least half of the mRNA molecules.
2. The composition of claim 1 , the mRNA molecules comprising a modified nucleotide.
3. The composition of claim 2 , the modified nucleotide comprising a modified pyrimidine.
4. The composition of claim 1 , the 5′ cap nucleoside being a 7-methylguanosine.
5. The composition of claim 2 , the 5′ cap nucleoside being a 7-methylguanosine.
6. The composition of claim 3 , the 5′ cap nucleoside being a 7-methylguanosine.
7. The composition of claim 1 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid comprising 1,2-distearoyl-sn-glycero-3-phosphocholine.
8. The composition of claim 2 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
9. The composition of claim 3 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
10. The composition of claim 4 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
11. The composition of claim 5 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
12. The composition of claim 6 , the lipid particles comprising the zwitterionic phospholipid; the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine; and at least 80% of the lipid particles having a diameter in the range of 20-220 nm.
13. The composition of claim 1 , the mRNA molecules being self-replicating RNA.
14. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 1 to elicit the immune response.
15. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 2 to elicit the immune response.
16. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 3 to elicit the immune response.
17. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 4 to elicit the immune response.
18. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 5 to elicit the immune response.
19. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 6 to elicit the immune response.
20. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 7 to elicit the immune response.
21. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 8 to elicit the immune response.
22. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 9 to elicit the immune response.
23. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 10 to elicit the immune response.
24. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 11 to elicit the immune response.
25. A method of eliciting in a human an immune response comprising an antibody response against the RSV F-protein immunogen or a cell-mediated immune response against the RSV F-protein immunogen, the method comprising administering to the human an effective amount of the composition of claim 24 to elicit the immune response.
26. The method of claim 14 comprising administering to the human at least two unit doses of the composition; the at least two unit doses being sequential and at least 1 week apart.
27. The method of claim 15 comprising administering to the human at least two unit doses of the composition; the at least two unit doses being sequential and at least 1 week apart.
28. The method of claim 16 comprising administering to the human at least two unit doses of the composition; the at least two unit doses being sequential and at least 1 week apart.
29. The method of claim 20 comprising administering to the human at least two unit doses of the composition; the at least two unit doses being sequential and at least 1 week apart.
30. The method of claim 25 comprising administering to the human at least two unit doses of the composition; the at least two unit doses being sequential and at least 1 week apart.