Infectious cDNA of an approved vaccine strain of measles virus, use for immunogenic compositions

摘要

The invention relates to a cDNA molecule which encodes the nucleotide sequence of the full length antigenomic (+)RNA strand of a measles virus (MV) originating from an approved vaccine strain. It also contains the preparation of immunogenic compositions using said cDNA.

主权项

1. 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.