Production of attenuated parainfluenza virus vaccines from cloned nucleotide sequences
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsBrian R. Murphy, Peter L. Collins, Anna P. Durbin, Mario H. Skiadopoulos, Tao Tao
Life of this patent
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Abstract
Isolated polynucleotide molecules provide recombinant PIV genomes and antigenomes for production of recombinant PIV vaccines. The recombinant genome or antigenome can be expressed with a nucleoprotein (N), phosphoprotein (P), and a large (L) polymerase protein to produce isolated infectious PIV particles. The recombinant PIV genome and antigenome can be modified to produce desired changes, for example to incorporate attenuating mutations from biologically derived PIV mutants or to create chimeric PIV clones, to generate attenuated, immunogenic viruses for vaccine use.
Claims70
An isolated polynucleotide molecule comprising an operably linked i) transcriptional promoter operative in a mammalian cell or operative in vitro, ii) a polynucleotide sequence encoding a partial or complete human PIV genome, said polynucleotide comprising at least one sequence selected from the group consisting of the complement of nucleotide sequences of SEQ ID NOS:
- 61, 63, 65, 67, 69, 71 and 73, or
- a polynucleotide encoding a partial or complete human PIV antigenome said polynucleotide comprising at least one nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 61, 63, 65, 67, 69, 71 and 73, and
- iii) a transcriptional terminator operative in a mammalian cell or operative in vitro.
An isolated polynucleotide molecule comprising an operably linked i) transcriptional promoter operative in a mammalian cell or operative in vitro, ii) a polynucleotide sequence encoding a partial or complete human PIV genome said polynucleotide comprising the complement of nucleotide sequences of SEQ ID NOS:
- 69, 71 and 73, or
- a polynucleotide encoding a partial or complete human PIV antigenome said polynucleotide comprising the nucleotide sequences of SEQ ID NOS:
- 69, 71 and 73, and
- iii) a transcriptional terminator operative in a mammalian cell or operative in vitro.
An isolated polynucleotide molecule comprising an operably linked transcriptional promoter operative in a mammalian cell or operative in vitro, a polynucleotide sequence encoding a chimeric partial or complete PIV genome or antigenome comprising said chimeric partial or complete partial or complete PIV genome or antigenome, and a transcriptional terminator operative in a mammalian cell or operative in vitro,
- wherein said polynucleotide sequence encoding said chimeric partial or complete PIV genome or antigenome comprises a polynucleotide sequence of a background partial or complete PIV genome or antigenome and at least one heterologous PIV sequence selected from a HPIV 1 sequence, a HPIV 2 sequence, a HPIV 3 sequence, a BPIV sequence or a MPIV sequence,
- and wherein said polynucleotide sequence encoding said chimeric partial or complete PIV genome or antigenome comprises a polynucleotide encoding the L protein of the wild-type of said background PIV or of said heterologous PIV.
An isolated polynucleotide molecule comprising an operably linked transcriptional promoter operative in a mammalian cell or operative in vitro, a polynucleotide sequence encoding a chimeric partial or complete PIV genome or antigenome that includes at least one point mutation comprising said partial or complete PIV genome or antigenome, and a transcriptional terminator operative in a mammalian cell or operative in vitro,
- wherein said polynucleotide sequence encoding said partial or complete PIV genome or antigenome comprises a polynucleotide sequence of a background partial or complete PIV genome or antigenome and at least one heterologous PIV sequence selected from a HPIV 1 sequence, a HPIV 2 sequence, a HPIV 3 sequence a BPIV sequence or a MPIV sequence,
- and wherein said polynucleotide sequence encoding said partial or complete PIV genome or antigenome comprises a polynucleotide encoding the L protein of the wild-type of said background PIV or of said heterologous PIV.
An isolated polynucleotide molecule comprising an operably linked transcriptional promoter operative in a mammalian cell or operative in vitro, a polynucleotide sequence encoding a chimeric partial or complete PIV genome or antigenome, and a transcriptional terminator operative in a mammalian cell or operative in vitro, wherein said polynucleotide sequence encoding said chimeric partial or complete PIV genome or antigenome comprises a polynucleotide sequence encoding a background partial or complete PIV genome or antigenome and at least one heterologous PIV polynucleotide sequence selected from a HPIV 1 sequence, a HPIV 2 sequence, a HPIV 3 sequence, a BPIV sequence or a MPIV sequence,
- wherein said heterologous PIV polynucleotide sequence comprises a transcription unit comprising a polynucleotide sequence encoding an open reading frame of a protein of said heterologous PIV or portion thereof providing an antigenic determinant that is inserted between a gene start sequence and a gene end sequence of the PIV of the background.
An infectious parainfluenza virus (PIV) comprising a major nucleocapsid (N) protein, a nucleocapsid phosphoprotein (P), a large polymerase protein (L), and a partial or complete human PIV genome comprising at least one sequence selected from the group consisting of the complement of nucleotide sequences of SEQ ID NOS:
- 61, 63, 65, 67, 69, 71 and 73,
- or a partial or complete human PIV antigenome comprising at least one nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 61, 63, 65, 67, 69, 71 and 73.
An infectious parainfluenza virus (PIV) comprising a major nucleocapsid (N) protein, a nucleocapsid phosphoprotein (P), a large polymerase protein (L), and a human partial or complete PIV genome comprising the complement of nucleotide sequences of SEQ ID NOS:
- 69, 71, and 73,
- or a partial or complete human PIV antigenome comprising the nucleotide sequences of SEQ ID NOS:
- 69, 71, and 73.
An infectious chimeric parainfluenza virus (PIV) comprising a major nucleocapsid (N) protein, a nucleocapsid phosphoprotein (P), a large polymerase protein (L), and a chimeric partial or complete PIV genome or antigenome comprising a a background partial or complete PIV genome or antigenome and at least one heterologous PIV sequence selected from a HPIV 1 sequence, a HPIV 2 sequence, a HPIV 3 sequence a BPIV sequence or a MPIV sequence,
- wherein said chimeric partial or complete PIV genome or antigenome comprises a polynucleotide encoding the L protein of the wild-type of said background PIV or of said heterologous PIV.
An infectious chimeric PIV virus comprising a chimeric partial or complete PIV genome or antigenome that includes at least one point mutation comprising said chimeric partial or complete PIV genome or antigenome,
- wherein said chimieric partial or complete PIV genome or antigenome comprises a polynucleotide sequence of a background partial or complete PIV genome or antigenome and at least one heterologous PIV sequence selected from a HPIV 1 sequence, a HPIV 2 sequence, a HPIV 3 sequence a BPIV sequence or a MPIV sequence,
- and wherein said polynucleotide sequence encoding said partial or complete PIV genome or antigenome comprises a polynucleotide encoding the L protein of the wild-type of said background PIV or of said heterologous PIV.
An infectious chimeric PIV comprising a chimeric partial or complete PIV genome or antigenome,
- wherein said chimeric partial or complete PIV genome or antigenome comprises a background partial or complete PIV genome or antigenome and at least one heterologous PIV polynucleotide sequence selected from a HPIV 1 sequence, a HPIV 2 sequence, a HPIV 3 sequence a BPIV sequence or a MPIV sequence,
- wherein said heterologous PIV polynucleotide sequence comprises a transcription unit comprising a polynucleotide sequence encoding an open reading frame of a protein of said heterologous PIV or portion thereof providing an antigenic determinant that is inserted between a gene start sequence and a gene end sequence of the PIV of the background.
Dependencies are inferred from the claim text.