The United States of America, As Represented by the Secretary, Department of Health and Human Services

The Salk Institute for Biological Studies

Michael Mackett

Bernard Moss

Geoffrey L Smith

Geoffrey Paul Smith

John K Rose

Tilahun Yilma

Recombinant poxviruses, such as vaccinia, are provided that comprises a segment comprised of (A) a first DNA sequence encoding a polypeptide that is foreign to poxvirus and (B) a poxvirus transcriptional regulatory sequence, wherein (i) said transcriptional regulatory sequence is adjacent to and exerts transcriptional control over said first DNA sequence and (ii) said segment is positioned within a nonessential genomic region of said recombinant poxvirus. Vaccines, carriers, cells, and media comprising recombinant poxviruses, and methods of immunization with recombinant poxviruses also are provided.

Methods of immunization using recombinant poxviruses having foreign DNA expressed under the control of poxvirus regulatory sequences

Methods and compositions are provided for the use of vaccinia virus or other poxviruses as vectors for expression of foreign genes. Expression of foreign genes is obtained by combining vaccinia virus transcriptional regulatory sequence with uninterrupted foreign protein coding sequences in vitro to form a chimeric gene. The chimeric gene is flanked by DNA from a non-essential region of the vaccinia virus genome to provide sites for in vivo homologous recombination. These steps are facilitated by the construction of plasmids that contain multiple restriction endonuclease sites, next to the vaccinia transcriptional regulatory sequences, for insertion of any foreign protein coding sequence. Transfection procedures are used to introduce the DNA into cells where homologous recombination results in the insertion of the chimeric gene into a non-essential region of the vaccinia virus genome. Infectious vaccinia virus recombinants are distinguished or selected by expression of the foreign gene, loss of activity of a vaccinia virus gene, or by DNA&#x2014;DNA hybridization. Expression of the foreign gene is obtained by infecting cells or animals with the recombinant vaccinia virus. Examples are provided to show expression of prokaryotic, RNA virus and other DNA virus genes in vaccinia recombinants.

Recombinant vaccinia virus containing a chimeric gene having foreign DNA flanked by vaccinia regulatory DNA

Recombinant poxviruses, such as vaccinia, are provided that comprises a segment comprised of (A) a first DNA sequence encoding a polypeptide that is foreign to poxvirus and (B) a poxvirus transcriptional regulatory sequence, wherein (i) said transcriptional regulatory sequence is adjacent to and exerts transcriptional control over said first DNA sequence and (ii) said segment is positioned within a nonessential genomic region of said recombinant poxvirus. Vaccines, carriers, cells, and media comprising recombinant poxviruses, and methods of immunization with recombinant poxviruses, also are provided.

Compositions containing recombinant poxviruses having foreign DNA expressed under the control of poxvirus regulatory sequences

Recombinant poxviruses having foreign DNA expressed under the control of poxvirus regulatory sequences

Synthetic vaccines for both the Indiana and New Jersey serotypes of vesicular stomatitis virus are provided. Recombinant vaccinia viruses are created with DNA sequences which encode antigenically active VSV proteins. Vaccinia virus sequences are inserted in plasmids and DNA sequences corresponding to encoding portions of the VSV genome are inserted into the plasmids with flanking vaccinia virus sequences. In cells transformed with such plasmids and also infected with vaccinia virus, homologous recombination occurs, producing the modified vaccinia viruses which act as vaccines encoding VSV proteins and inducing anti-VSV immune responses in inoculated animals.

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Michael Mackett