Immunisation of large mammals with low doses of RNA
Provided is RNA encoding an immunogen is delivered to a large mammal at a dose of from 5 μg to 100 μg or at a dose between 0.1 μg per kilogram body mass of the large mammal and 1.5 μg per kilogram body mass of the large mammal. Provided is a method of raising an immune response in a large mammal, comprising administering to the large mammal a dose of from 5 μg to 100 μg or a dose of between 0.1 μg of RNA encoding the immunogen per kilogram body mass of the large mammal and 1.5 μg of RNA encoding the immunogen per kilogram body mass of the large mammal. 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 intramuscularly to the large mammal at least two unit doses; each unit dose comprising a composition comprising lipid particles and messenger ribonucleic acid (mRNA) molecules; the at least two unit doses being sequential and administered at least 1 week apart; the administering comprising contacting the composition with skeletal muscle; the mRNA molecules comprising 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, 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 lipid particles comprising: i) a polyethylene glycol-ylated lipid, ii) cholesterol, iii) an anionic phospholipid or a zwitterionic phospholipid, and iv) a cationic lipid; the cationic lipid comprising a tertiary amine; the lipid particles encapsulating at least half of the mRNA molecules; and the large mammal being a human, a horse, a cow, or a pig.
2. A method of eliciting an antibody response against an immunogen by an immune system in a large mammal, the method comprising administering intramuscularly to the large mammal at least two unit doses; each unit dose comprising a composition comprising lipid particles and mRNA molecules; the at least two unit doses being sequential and administered at least 1 week apart; the administering comprising contacting the composition with skeletal muscle; the mRNA molecules comprising a sequence that encodes the immunogen; the immunogen comprising a RSV F protein immunogen, an EBV glycoprotein immunogen, a CMV glycoprotein immunogen, a HPV 16 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 lipid particles comprising: i) a polyethylene glycol-ylated lipid, ii) cholesterol, iii) an anionic phospholipid or a zwitterionic phospholipid, and iv) a cationic lipid; the cationic lipid comprising a tertiary amine; the lipid particles encapsulating at least half of the mRNA molecules; and the large mammal being a human, a horse, a cow, or a pig.
3. The method of claim 1 , the mRNA molecules comprising a 5′ cap nucleoside, a first 5′ ribonucleoside, and a triphosphate bride; the 5′ cap nucleoside being linked 5′-to-5′ to the first 5′ ribonucleoside by the triphosphate bridge; the 5′ cap nucleoside optionally being a 7-methylguanosine; and the large mammal being the human.
4. The method of claim 2 , the mRNA molecules comprising a 5′ cap nucleoside, a first 5′ ribonucleoside, and a triphosphate bride; the 5′ cap nucleoside being linked 5′-to-5′ to the first 5′ ribonucleoside by the triphosphate bridge; the 5′ cap nucleoside optionally being a 7-methylguanosine; and the large mammal being the human.
5. The method of claim 3 , at least 80% of the lipid particles having diameters from 20 nm to 220 nm.
6. The method of claim 4 , at least 80% of the lipid particles having diameters from 20 nm to 220 nm.
7. The method of claim 3 , the mRNA molecules comprising a modified nucleotide.
8. The method of claim 4 , the mRNA molecules comprising a modified nucleotide.
9. The method of claim 7 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1 ,2-distearoyl-sn-glycero-3-phosphocholine.
10. The method of claim 8 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1 ,2-distearoyl-sn-glycero-3-phosphocholine.
11. The method of claim 7 , the modified nucleotide being a modified pyrimidine nucleotide.
12. The method of claim 8 , the modified nucleotide being a modified pyrimidine nucleotide.
13. The method of claim 7 , the immunogen comprising the flavivirus immunogen.
14. The method of claim 8 , the immunogen comprising the flavivirus immunogen.
15. The method of claim 7 , comprising administering to the human at least three unit doses.
16. The method of claim 8 , comprising administering to the human at least three unit doses.
17. The method of claim 9 , comprising administering to the human at least three unit doses.
18. The method of claim 10 , comprising administering to the human at least three unit doses.
19. The method of claim 1 , further eliciting a cell-mediated immune response by the immune system in the large mammal.
20. The method of claim 2 , further eliciting a cell-mediated immune response by the immune system in the large mammal.
21. The method of claim 3 , further eliciting a cell-mediated immune response by the immune system in the human.
22. The method of claim 4 , further eliciting a cell-mediated immune response by the immune system in the human.
23. The method of claim 5 , further eliciting a cell-mediated immune response by the immune system in the human.
24. The method of claim 6 , further eliciting a cell-mediated immune response by the immune system in the human.
25. The method of claim 7 , further eliciting a cell-mediated immune response by the immune system in the human.
26. The method of claim 8 , further eliciting a cell-mediated immune response by the immune system in the human.
27. The method of claim 9 , further eliciting a cell-mediated immune response by the immune system in the human.
28. The method of claim 10 , further eliciting a cell-mediated immune response by the immune system in the human.
29. The method of claim 13 , further eliciting a cell-mediated immune response by the immune system in the human.
30. The method of claim 14 , further eliciting a cell-mediated immune response by the immune system in the human.
31. The method of claim 15 , further eliciting a cell-mediated immune response by the immune system in the human.
32. The method of claim 16 , further eliciting a cell-mediated immune response by the immune system in the human.
33. The method of claim 17 , further eliciting a cell-mediated immune response by the immune system in the human.
34. The method of claim 18 , further eliciting a cell-mediated immune response by the immune system in the human.
35. The method of claim 1 , the mRNA molecules being self-replicating RNA molecules.
36. The method of claim 2 , the mRNA molecules being self-replicating RNA molecules.