Parainfluenza virus 5 based vaccines
The present invention provides safe, stable, efficacious, and cost-effective vaccines based on viral expression vectors that include a parainfluenza virus 5 (PIV5) genome including a heterologous nucleotide sequence expressing a heterologous polypeptide. In some embodiments, the heterologous nucleotide sequence is inserted closer to the leader than between the hemagglutinin-neuroaminidase (HN) gene and the large RNA polymerase protein (L) gene of the PIV5 genome. In some embodiments, the heterologous nucleotide sequence is inserted between the small hydrophobic protein (SH) gene and the hemagglutinin-neuroaminidase (HN) gene of the PIV 5 genome.
1. A viral expression vector comprising a parainfluenza virus 5 (PIV5) genome comprising a heterologous nucleotide sequence expressing a heterologous polypeptide, wherein the heterologous nucleotide sequence is inserted between the small hydrophobic protein (SH) gene and the hemagglutinin-neuraminidase (HN) gene of the PIV5 genome.
2. The viral expression vector of claim 1 , wherein the PIV5 genome further comprises one or more mutations.
3. The viral expression vector of claim 2 , wherein a mutation comprises a mutation of the V/P gene, a mutation of the shared N-terminus of the V and P proteins, a mutation of residues 26, 32, 33, 50, 102, and/or 157 of the shared N-terminus of the V and P proteins, a mutation lacking the C-terminus of the V protein, a mutation lacking the small hydrophobic (SH) protein, a mutation of the fusion (F) protein, a mutation of the phosphoprotein (P), a mutation of the large RNA polymerase (L) protein, a mutation incorporating residues from canine parainfluenza virus, a mutation inducing apoptosis, or a combination thereof.
4. The viral expression vector of claim 2 , wherein the mutation comprises PIV5VΔC, PIV5ΔSH, PIV5-P-S308G, or a combination thereof.
5. The viral expression vector of claim 1 , wherein the heterologous polypeptide comprises an influenza polypeptide.
6. The viral expression vector of claim 5 , wherein the influenza comprises influenza A, influenza B, or influenza C virus.
7. The viral expression vector of claim 5 , wherein the heterologous polypeptide comprises a hemagglutinin (HA) from influenza A virus strain subtype H1 to H18.
8. The viral expression vector of claim 5 , wherein the heterologous polypeptide comprises a hemagglutinin (HA) from influenza A virus strain H5N1, H3N2, or H1N1.
9. The viral expression vector of claim 5 , wherein the heterologous polypeptide comprises an influenza neuraminidase (NA) from influenza type A subtype N1 to N10.
10. The viral expression vector of claim 5 , wherein the influenza comprises influenza A virus strain H5N1, H3N2, or H1N1.
11. The viral expression vector of claim 5 , wherein the heterologous influenza polypeptide comprises an influenza hemagglutinin (HA), an influenza neuraminidase (NA), an influenza nucleocapsid protein (NP), M1, M2, PA, PB1, PB2, PB1-F2, NS1 or NS2.
12. The viral expression vector of claim 1 , wherein the heterologous polypeptide is derived from human immunodeficiency virus (HIV), parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, human respiratory syncytial virus, bovine respiratory syncytial virus, human metapneumovirus, avian influenza, canine influenza, avian metapneumovirus, Nipah virus, Hendra virus, rabies virus, Ebola virus, porcine circovirus, porcine reproductive and respiratory syndrome virus, swine influenza virus, New Castle disease virus (NDV), Marek's disease virus (MDV), infectious bronchitis virus (IBV), egg drop syndrome (EDS) virus, turkey rhinotracheitis virus (TRTV), mumps virus, measles virus, canine distemper virus, feline leukemia virus, human calicivirus, veterinary calcivirus, feline calcivirus, human norovirus, veterinary norovirus, rinderpest virus, and/or Mycobacterium tuberculosis.
13. The viral expression vector of claim 1 comprising two or more heterologous nucleotide sequence expressing a heterologous polypeptide.
14. A viral particle comprising a viral expression vector of claim 1 .
15. A composition of the viral expression vector of claim 1 or a viral particle comprising the viral expression vector of claim 1 .
16. The viral expression vector of claim 1 , wherein the heterologous polypeptide comprises a rabies virus nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G), and/or viral RNA polymerase (L).
17. The viral expression vector of claim 1 , wherein the heterologous polypeptide comprises a respiratory syndrome virus (RSV) F protein and/or or RSV G protein.
18. The viral expression vector of claim 1 , wherein the heterologous polypeptide comprises a Mycobacterium tuberculosis 85A, 85B and/or ESAT6 protein.
19. The viral expression vector of claim 1 , wherein the heterologous polypeptide comprises a human immunodeficiency virus (HIV) gp160 antigen, gp140 antigen, gp120 antigen, and/or Gag antigen.
20. The viral expression vector of claim 1 , wherein the heterologous polypeptide comprises a hemagglutinin (HA), neuraminidase (NA), and/or nucleocapsid protein (NP) of influenza strain H5N1.
21. The viral expression vector of claim 1 further comprising a second heterologous nucleotide sequence expressing a second heterologous polypeptide inserted between the hemagglutinin-neuroaminidase (HN) gene and the large RNA polymerase protein (L) gene of the PIV5 genome.
22. A method of expressing a heterologous polypeptide in a cell, the method comprising contacting the cell with a viral expression vector, viral particle, or composition of claim 1 .
23. A method of inducing an immune response in a subject to a heterologous polypeptide, the method comprising administering a viral expression vector, viral particle, or composition of claim 1 to the subject.
24. The method of claim 23 comprising an avian subject.
25. The method of claim 24 , comprising in ovo administration.
26. The method of claim 23 , wherein the subject demonstrates pre-existing immunity to PIV5.