Attenuated vaccines for non-segmented negative sense rna viruses

Abstract

The invention relates to an attenuated non-segmented negative-sense RNA virus characterized by at least one mutation in the L gene wherein the mutation reduces viral replication, the methods of manufacturing and methods of use.

1 .- 27 . (canceled) 
     
     
         28 . An isolated attenuated non-segmented negative-sense RNA virus comprising one or more mutations in Domain VI of the L gene, wherein the one or more mutations result in a decrease in mRNA cap methylation. 
     
     
         29 . The virus of  claim 28 , wherein the one or more mutations comprise substitutions of amino acids selected from the group consisting of K1651, G1670, G1672, S1673, G1675, D1735, D1762, K1795, and E1833 of vesicular stomatitis virus (VSV) or the corresponding amino acids of other non-segmented negative-sense RNA viruses. 
     
     
         30 . The virus of  claim 29 , further comprising a substitution of amino acid D1671 of vesicular stomatitis virus (VSV) or the corresponding amino acid of other non-segmented negative-sense RNA viruses. 
     
     
         31 . The virus of  claim 28 , wherein the virus comprises at least two, at least three, or at least four mutations. 
     
     
         32 . The virus of  claim 28  wherein the one or more mutations results in the substitution of an amino aid located on the L protein surface as predicted by protein modeling or crystallography. 
     
     
         33 . The virus of  claim 28  wherein mRNA cap methylation activity is reduced by at least 50%. 
     
     
         34 . The virus of  claim 28  wherein the virus substantially retains its capacity to express N, P, M, or G proteins. 
     
     
         35 . The virus of  claim 34 , wherein expression of N, P, M, or G proteins comprises at least 30%, at least 40%, at least 50%, or at least 60% of wild-type expression levels. 
     
     
         36 . The virus of  claim 28  further comprising one or more heterologous polynucleotides. 
     
     
         37 . The virus according to  claim 36  wherein the one or more heterologous polynucleotides encode one or more antigens. 
     
     
         38 . The virus of  claim 37 , wherein the one or more antigens comprise antigens from one or more pathogens. 
     
     
         39 . The virus of  claim 38 , wherein the one or more pathogens comprise viruses, bacteria, fungi, or parasites. 
     
     
         40 . The virus of  claim 38 , wherein the one or more pathogens are selected from a group consisting of HIV, HTLV, mycobacteria, influenza, respiratory syncytial virus, and hepatitis B virus. 
     
     
         41 . The virus of  claim 37 , wherein the one or more antigens comprise tumor antigens. 
     
     
         42 . The virus of  claim 36 , wherein the one or more heterologous polynucleotides comprise RNA, siRNA, or microRNA. 
     
     
         43 . A pharmaceutical composition comprising the virus of  claim 28  and a pharmaceutically acceptable carrier. 
     
     
         44 . A method of vaccinating a human or animal subject, comprising administering to the subject an effective amount of the pharmaceutical composition of  claim 42 . 
     
     
         45 . A method of treating a tumor in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of  claim 42 . 
     
     
         46 . The virus of  claim 28 , wherein the virus is of the order of Mononegavirales or the family Rhabdoviridae. 
     
     
         47 . The virus of  claim 28 , wherein the virus is vesicular stomatitis virus.