Synergistic attenuation of vesicular stomatitis virus, vectors thereof and immunogenic compositions thereof

1. A genetically modified vesicular stomatitis virus (VSV) comprising at least two different classes of mutations in the same VSV genome, wherein the VSV comprises a truncated G gene mutation and an N gene shuffling mutation, wherein the two mutations synergistically attenuate VSV pathogenicity.

2. The VSV of claim 1 , wherein the pathogenicity is further defined as neurovirulence.

3. The VSV of claim 1 , wherein the G protein encoded by the truncated G gene has a deletion of the last twenty or twenty-eight carboxy-terminal amino acids.

4. The VSV of claim 1 , wherein the G protein encoded by the truncated G gene has a cytoplasmic tail domain consisting of one or nine amino acids.

5. The VSV of claim 1 , wherein the N gene is shuffled to 3′-PNMGL-5′ or 3′-PMNGL-5′, relative to the wild-type VSV genome 3′-NP GL-5′, wherein N is the gene encoding the nucleocapsid protein, P is the gene encoding the phosphoprotein, M is the gene encoding the matrix protein, G is the gene encoding the attachment glycoprotein and L is the gene encoding the RNA-dependent RNA polymerase protein.

6. The VSV of claim 1 , comprising a mutated genome selected from the group consisting of 3′-PNMG (ct-1) L-5′, 3′-PNMG (ct-9) L-5′, 3′-PMNG (ct-1) L-5′ and 3′-PMNG (ct-9) L-5′, wherein N is the gene encoding the nucleocapsid protein. P is the gene encoding the phosphoprotein, M is the gene encoding the matrix protein. G (ct-1) is the gene encoding the attachment glycoprotein having a deletion of the last twenty-eight carboxy-terminal amino acids, G (ct-9) is the gene encoding the attachment glycoprotein having a deletion of the last twenty carboxy-terminal amino acids and L is the gene encoding the RNA-dependent RNA polymerase protein.

7. The VSV of claim 6 , further comprising a third class of mutation in the same VSV genome, wherein the mutation is a ts mutation, an ambisense RNA mutation, a non-cytopathic M gene mutation, a gene deletion mutation, a gene insertion mutation or a point mutation.

8. The VSV of claim 1 , wherein the VSV injected intracranially in 4-week old female Swiss-Webster mice has a LD 50 100-fold, 1,000-fold, 10,000-fold, or 100,000-fold greater than wild-type VSV injected intracranially in 4-week old female Swiss-Webster mice.

9. The VSV of claim 1 , wherein the N gene shuffling mutation results from an insertion of at least one foreign RNA sequence into the VSV genome at position 1.

10. A genetically modified VSV vector comprising the VSV of claim 1 , further comprising at least one foreign RNA sequence inserted into or replacing a region of the VSV genome non-essential for replication.

11. The vector of claim 10 , wherein the foreign RNA is further defined as an open reading frame (ORF).

12. The vector of claim 10 , wherein the foreign RNA is selected from the group consisting of an HIV gene, an HTLV gene, an SIV gene, an RSV gene, a PIV gene, an HSV gene, a CMV gene, an Epstein-Barr virus gene, a Varicella-Zoster virus gene, a mumps virus gene, a measles virus gene, an influenza virus gene, a poliovirus gene, a rhinovirus gene, a hepatitis A virus gene, a hepatitis B virus gene, a hepatitis C virus gene, a Norwalk virus gene, a togavirus gene, an alphavirus gene, a rubella virus gene, a rabies virus gene, a Marburg virus gene, an Ebola virus gene, a papilloma virus gene, a polyoma virus gene, a metapneumovirus gene, a coronavirus gene, a Vibrio cholerae gene, a Streptococcus pneumoniae gene, a Streptococcus pyogenes gene, a Helicobacter pylori gene, a Streptococcus agalactiae gene, a Neisseria meningitidis gene, a Neisseria gonorrheae gene, a Corynebacteria diphtheriae gene, a Clostridium tetani gene, a Bordetella pertussis gene, a Haemophilus gene, a Chlamydia gene, an Escherichia coli gene, a gene encoding a cytokine, a gene encoding T-helper epitope, a gene encoding a CTL epitope, a gene encoding an adjuvant and a gene encoding a co-factor.

13. The vector of claim 12 , wherein the foreign RNA is an HIV gene selected from the group consisting of gag, env, pol, vif, nef, tat, vpr, rev and vpu.

14. The vector of claim 13 , wherein the HIV gene is gag and the mutated genome is 3′-gag 1 -PNMG (ct-1) L-5′, 3′-gag 1 -PNMG (ct-9) )L-5′, 3′-gag 1 -PMNG (ct-9) L-5′, 3′-gag 1 -PMNG (ct-9) L-5′, 3′-PNMG (ct-1) -gag 5 -L-5′, 3′-PMNG (ct-9) -gag 5 -L-5′ or 3′-PMNG (ct-9) -gag 5 -L-5′.

15. An immunogenic composition comprising an immunogenic dose of the VSV vector of claim 10 .

16. The VSV of claim 9 , wherein the foreign RNA sequence is an HIV gene.

17. The VSV of claim 16 , wherein the HIV gene is gag.