Vaccines against infectious diseases caused by positive stranded rna viruses

Abstract

Compositions for protecting subjects from diseases caused by (+) SS RNA virus are described herein. The compositions include (i) a vector containing a DNA encoding a RNA molecule of an infectious (+) SS RNA virus operably linked to a eukaryotic RNA polymerase promoter and a carrier; or (ii) (+) SS RNA viruses obtained from eukaryotic cells transfected with the vector of (i) and a carrier.

1 . A homogeneous population of clonally purified positive single stranded ((+)SS) RNA virus obtained from eukaryotic cells transfected with a vector, wherein the vector comprises a DNA encoding a RNA molecule operably linked to a promoter suitable for expression of the DNA in a eukaryotic cell, wherein the RNA molecule encodes an infectious (+)SS RNA virus, wherein the (+)SS) RNA virus is from the family Flaviviridae, wherein the DNA comprises at least three introns, and wherein
 the DNA encoding the RNA molecule comprises SEQ ID NO:1;   the DNA encoding the RNA molecule comprises at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 1, and the introns in SEQ ID NO: 1 are intact;   the DNA comprises at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity with a DNA encoding nucleic acid sequence of JEV SA14-14-2 strain and the introns are inserted immediately after nucleotide 414, nucleotide 2213, and nucleotide 3134 of the DNA encoding the nucleic acid sequence of JEV SA14-14-2 strain; or   the DNA encodes nucleic acid sequence of JEV SA14-14-2 strain and the three introns are inserted immediately after nucleotide 414, nucleotide 2213, and nucleotide 3134 of the DNA encoding JEV SA14-14-2 strain.   
     
     
         2 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 1 , wherein the (+)SS RNA virus comprises a flavivirus. 
     
     
         3 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 2 , wherein the flavivirus is JEV. 
     
     
         4 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 1 , wherein at least one of the introns contain a stop codon or several stop codons. 
     
     
         5 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 1 , wherein the clonally purified (+)SS RNA virus is a nonpathogenic virus and/or an attenuated virus. 
     
     
         6 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 1 , wherein the promoter comprises a CMV promoter, a RSV promoter, a SV40 promoter, a HSV promoter, a human Pol I promoter, a human Pol II promoter, or a human Pol III promoter. 
     
     
         7 . A composition comprising the homogeneous population of clonally purified (+)SS RNA virus of  claim 1  and a carrier. 
     
     
         8 . An immunogenic composition comprising an effective amount of the homogeneous population of clonally purified (+) SS RNA virus of  claim 1  and a carrier. 
     
     
         9 . A pharmaceutical composition comprising an effective amount of the homogeneous population of clonally purified (+)SS RNA virus of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         10 . An immunogenic composition comprising an effective amount of the homogeneous population of clonally purified live (+)SS RNA virus of  claim 5 . 
     
     
         11 . A homogeneous population of clonally purified (+)SS RNA virus obtained from eukaryotic cells transfected with a vector, wherein the vector comprises a DNA encoding a chimeric RNA molecule of a chimeric infectious (+)SS RNA virus, wherein the DNA is operably linked to a promoter suitable for expression of the DNA in a eukaryotic cell,
 wherein the chimeric RNA molecule comprises nucleic acid sequences from at least two different (+)SS RNA viruses from the family Flaviviridae, wherein the DNA comprises at least three introns, and wherein
 the DNA encoding the chimeric RNA molecule comprises nucleotides 1001 to 4477 of SEQ ID NO: 1; and 
 the DNA encoding the chimeric RNA comprises at least 80% or 85% of the entire nucleic acid sequence of JEV SA14-14-2 strain. 
   
     
     
         12 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 11 , wherein the (+)SS RNA virus is encoded by a chimeric RNA molecule comprising the RNA of at least two different flaviviruses. 
     
     
         13 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 12 , wherein the at least two different flaviviruses comprise JEV and at least one of Dengue virus, Yellow fever virus, West Nile virus, tick borne encephalitis virus, Hepatitis C virus, or Zika virus. 
     
     
         14 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 13 , wherein the at least two different flaviviruses are JEV and Zika virus. 
     
     
         15 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 11 , wherein at least one of the introns contain a stop codon or several stop codons. 
     
     
         16 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 11 , wherein the clonally purified (+)SS RNA virus is a nonpathogenic virus and/or an attenuated virus. 
     
     
         17 . The homogeneous population of clonally purified (+)SS RNA virus of  claim 11 , wherein the promoter comprises a CMV promoter, a RSV promoter, a SV40 promoter, a HSV promoter, a human Pol I promoter, a human Pol II promoter, or a human Pol III promoter. 
     
     
         18 . A composition comprising the homogeneous population of clonally purified (+)SS RNA virus of  claim 11  and a carrier. 
     
     
         19 . An immunogenic composition comprising an effective amount of the homogeneous population of clonally purified (+)SS RNA virus of  claim 11  and a carrier. 
     
     
         20 . A pharmaceutical composition comprising an effective amount of the homogeneous population of clonally purified (+)SS RNA virus of  claim 11  and a pharmaceutically acceptable carrier. 
     
     
         21 . An immunogenic composition comprising an effective amount of the homogeneous population of clonally purified live (+)SS RNA virus of  claim 16 . 
     
     
         22 . A method of protecting a subject against a disease caused by a (+)SS RNA virus, wherein the method comprises administering the homogeneous population of clonally purified (+) SS RNA virus of  claim 1  to a subject in need thereof, and wherein the disease is caused by flavivirus. 
     
     
         23 . The method of  claim 22 , wherein the flavivirus comprises JEV. 
     
     
         24 . A method of protecting a subject against a disease caused by a (+)SS RNA virus, wherein the method comprises administering the homogeneous population of clonally purified (+)SS RNA virus of  claim 5  to a subject in need thereof, and wherein the disease is caused by flavivirus. 
     
     
         25 . The method of  claim 24 , wherein the flavivirus comprises JEV. 
     
     
         26 . The method of  claim 24 , wherein the (+)SS RNA virus is encoded by a chimeric RNA molecule comprising the RNA of at least two different flaviviruses. 
     
     
         27 . The method of  claim 26 , wherein the at least two different flaviviruses comprise JEV and at least one of Dengue virus, Yellow fever virus, West Nile virus, tick borne encephalitis virus, Hepatitis C virus, or Zika virus.