Immunization 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 a respiratory syncytial virus (RSV) surface fusion glycoprotein (F protein) 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 RSV F protein 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 or a cow.
2. The method of claim 1 , the mRNA molecules comprising a 5′ cap nucleoside, a first 5′ ribonucleoside, and a triphosphate bridge; 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.
3. The method of claim 2 , the mRNA molecules comprising a modified nucleotide.
4. The method of claim 3 , the modified nucleotide being a modified pyrimidine nucleotide.
5. The method of claim 4 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
6. The method of claim 5 comprising administering to the human at least three unit doses.
7. The method of claim 3 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
8. The method of claim 7 comprising administering to the human at least three unit doses.
9. The method of claim 2 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
10. The method of claim 9 comprising administering to the human at least three unit doses.
11. The method of claim 1 , at least 80% of the lipid particles having diameters from 20 nm to 220 nm.
12. The method of claim 11 , the lipid particles comprising the zwitterionic phospholipid; the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine; and the large mammal being the human.
13. The method of claim 12 comprising administering to the human at least three unit doses.
14. The method of claim 1 , the lipid particles comprising the zwitterionic phospholipid; the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine; and the large mammal being the human.
15. The method of claim 14 comprising administering to the human at least three unit doses.
16. A method of eliciting an antibody response against a RSV F protein 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 RSV F protein 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 or a cow.
17. The method of claim 16 , the mRNA molecules comprising a 5′ cap nucleoside, a first 5′ ribonucleoside, and a triphosphate bridge; 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.
18. The method of claim 17 , the mRNA molecules comprising a modified nucleotide i.
19. The method of claim 18 , the modified nucleotide being a modified pyrimidine nucleotide.
20. The method of claim 19 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
21. The method of claim 20 comprising administering to the human at least three unit doses.
22. The method of claim 18 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
23. The method of claim 22 comprising administering to the human at least three unit doses.
24. The method of claim 17 , the lipid particles comprising the zwitterionic phospholipid; and the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine.
25. The method of claim 24 comprising administering to the human at least three unit doses.
26. The method of claim 16 , at least 80% of the lipid particles having diameters from 20 nm to 220 nm.
27. The method of claim 26 , the lipid particles comprising the zwitterionic phospholipid; the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine; and the large mammal being the human.
28. The method of claim 27 comprising administering to the human at least three unit doses.
29. The method of claim 16 , the lipid particles comprising the zwitterionic phospholipid; the zwitterionic phospholipid being 1,2-distearoyl-sn-glycero-3-phosphocholine; and the large mammal being the human.
30. The method of claim 29 comprising administering to the human at least three unit doses.