Recombinant measles viruses expressing epitopes of antigens of rna viruses - use for the preparation of vaccine compositions

Abstract

The invention relates to a recombinant measles virus expressing a heterologous amino acid sequence derived from an antigen of a determined RNA virus, said recombinant measles virus being capable of eliciting a humoral and/or cellular immune response against measles virus or against said RNA virus or against both measles virus and against said RNA virus. It also relates to the use of said recombinant measles virus for the preparation of immunogenic composition.

1 - 51 . (canceled) 
     
     
         52 . A method of inducing an immune response against an RNA virus in a host, comprising administering to the host a composition comprising (i) a recombinant measles virus expressing a heterologous amino acid sequence, or a recombinant measles virus expression vector for expressing a heterologous amino acid sequence, and (ii) an acceptable vehicle, wherein said virus or vector comprises a sequence comprising:
 A) a nucleotide sequence encoding the full length antigenomic (+)RNA strand of a measles virus;   B) a T7 promoter sequence comprising a GGG motif at its 3′ end, operably linked to the nucleotide sequence of A;   C) a hammerhead ribozyme sequence located adjacent to the GGG motif at one end and adjacent to the first nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus strain at the other end;   D) a T7 terminator sequence operably linked to the nucleotide sequence of A;   E) the sequence of a hepatitis delta virus ribozyme located adjacent to the last nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus; and   F) a heterologous coding sequence encoding a heterologous amino acid sequence comprising an antigen of a heterologous RNA virus.   
     
     
         53 . The method of  claim 52 , wherein the measles virus is a measles virus vaccine strain. 
     
     
         54 . The method of  claim 52 , wherein the RNA virus is a flavivirus or a retrovirus. 
     
     
         55 . The method of  claim 52 , wherein the RNA virus is not a flavivirus or a retrovirus. 
     
     
         56 . The method of  claim 52  wherein the heterologous coding sequence is cloned between the P and M genes of the measles virus. 
     
     
         57 . The method according to  claim 52 , wherein the heterologous coding sequence is cloned between the H and L genes of the measles virus. 
     
     
         58 . The method according to  claim 52 , wherein the composition is administered in a prime-boost administration regime. 
     
     
         59 . The composition according to  claim 52 , wherein the composition is administered through subcutaneous (s.c.) or intraperitoneal (i.p.) injection. 
     
     
         60 . The method of  claim 53 , wherein the RNA virus is not a flavivirus or a retrovirus. 
     
     
         61 . The method of  claim 60  wherein the heterologous coding sequence is cloned between the P and M genes of the measles virus. 
     
     
         62 . The method according to  claim 60 , wherein the heterologous coding sequence is cloned between the H and L genes of the measles virus. 
     
     
         63 . The method according to  claim 60 , wherein the composition is administered in a prime-boost administration regime. 
     
     
         64 . The composition according to  claim 60 , wherein the composition is administered through subcutaneous (s.c.) or intraperitoneal (i.p.) injection. 
     
     
         65 . An expression vector for producing an infectious recombinant Schwarz strain of measles virus comprising:
 A) the nucleotide sequence encoding the full length antigenomic (+)RNA strand of the measles virus Schwarz strain (from position 83 to position 15976 of SEQ ID NO: 82);   B) a T7 promoter sequence comprising a GGG motif at its 3′ end, operably linked to the nucleotide sequence of A;   C) a hammerhead ribozyme sequence (from position 29 to position 82 of SEQ ID NO: 82) located adjacent to the GGG motif at one end and adjacent to the first nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus Schwarz strain at the other end;   D) a T7 terminator sequence operably linked to the nucleotide sequence of A;   E) the sequence of a hepatitis delta virus ribozyme located adjacent to the last nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus Schwarz strain; and   F) a heterologous coding sequence encoding a heterologous amino acid sequence.   
     
     
         66 . A method for generating infectious recombinant Schwarz strain of measles virus, comprising:
 (A) providing an expression vector for producing the Schwarz strain of measles virus, wherein the expression vector comprises
 (i) the nucleotide sequence encoding the full length antigenomic (+)RNA strand of the measles virus Schwarz strain (from position 83 to position 15976 of SEQ ID NO: 82); 
 (ii) a T7 promoter sequence comprising a GGG motif at its 3′ end, operably linked to the nucleotide sequence of (i); 
 (iii) a hammerhead ribozyme sequence (from position 29 to position 82 of SEQ ID NO: 82) located adjacent to the GGG motif at one end and adjacent to the nucleotide sequence of (i) at the other end; 
 (iv) a T7 terminator sequence operably linked to the nucleotide sequence of A; 
 (v) the sequence of a hepatitis delta virus ribozyme located adjacent to the last nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus Schwarz strain; 
 (vi) a heterologous coding sequence encoding an amino acid sequence of a heterologous antigen; 
   (B) transfecting helper cells with the expression vector of (A), wherein the helper cells express a heterologous DNA-dependent RNA polymerase and N, P, and L proteins of measles virus;   (C) maintaining the transfected helper cells under conditions suitable for production of Schwarz strain measles viral particles;   (D) passaging the viral particles in cells suitable for the passage of the Schwarz strain; and   (E) recovering infectious recombinant Schwarz strain measles virus particles.   
     
     
         67 . An expression vector for producing an infectious recombinant live-attenuated measles virus comprising:
 A) a nucleotide sequence encoding a full length antigenomic (+)RNA strand of the live-attenuated measles virus;   B) a T7 promoter sequence comprising a GGG motif at its 3′ end, operably linked to the nucleotide sequence of A);   C) a hammerhead ribozyme sequence located adjacent to the GGG motif at one end and adjacent to the first nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus at the other end;   D) a T7 terminator sequence operably linked to the nucleotide sequence of A);   E) a sequence of a hepatitis delta virus ribozyme located adjacent to the last nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus; and   F) a heterologous coding sequence encoding a heterologous amino acid sequence.   
     
     
         68 . A method of making an immunogenic composition, comprising:
 providing (i) a recombinant measles virus expressing a heterologous amino acid sequence, or a recombinant measles virus expression vector for expressing a heterologous amino acid sequence, wherein said virus or vector comprises a sequence comprising:   A) a nucleotide sequence encoding the full length antigenomic (+)RNA strand of a measles virus;   B) a T7 promoter sequence comprising a GGG motif at its 3′ end, operably linked to the nucleotide sequence of A;   C) a hammerhead ribozyme sequence located adjacent to the GGG motif at one end and adjacent to the first nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus strain at the other end;   D) a T7 terminator sequence operably linked to the nucleotide sequence of A;   E) the sequence of a hepatitis delta virus ribozyme located adjacent to the last nucleotide of the nucleotide sequence encoding the full length anti-genomic (+)RNA strand of the measles virus; and   F) a heterologous coding sequence encoding a heterologous amino acid sequence comprising an antigen of a heterologous RNA virus; and   combining the recombinant measles virus expressing a heterologous amino acid sequence, or a recombinant measles virus expression vector for expressing a heterologous amino acid sequence, with (ii) an acceptable vehicle, to thereby provide the immunogenic composition.