Recombinant BCG
AssigneesBeth Israel Hospital, Whitehead Institute for Biomedical Research
InventorsMichael A. O'Donnell, Rosemary B. Duda, William C. DeWolf, Anna Aldovini, Richard A. Young
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Abstract
The present invention relates to recombinant mycobacteria, particularly recombinant M. bovis BCG, which express heterologous DNA encoding a product (protein or polypeptide) of interest, such a protein or polypeptide (e.g., an antigen) against which an immune response is desired or a cytokine.
Claims28
A recombinant mycobacterium having enhanced immunostimulatory properties in comparison with immunostimulatory properties of wild type mycobacterium, comprising DNA encoding a cytokine, wherein the DNA is expressed extrachromosomally under the control of a mycobacterial heat shock protein gene promoter or a mycobacterial stress protein gene promoter and the cytokine is secreted from said mycobacterium in a biologically active form.
Recombinant BCG having enhanced immunostimulatory properties in comparison with immunostimulatory properties of wild type BCG and having incorporated therein a plasmid comprising DNA encoding a cytokine operably linked to a mycobacterial heat shock protein gene promoter and a mycobacterial secretion signal sequence wherein the 5' to 3' order of the plasmid is the promoter, the secretion signal sequence and the DNA encoding a cytokine, and the cytokine is expressed and secreted from the BCG in a biologically active form.
A mycobacterial autonomous replicating plasmid vector which, when expressed in mycobacteria, results in enhanced immunostimulatory properties of the mycobacteria, in comparison with immunostimulatory properties of mycobacteria in which the vector is not expressed, the plasmid vector comprising:
- a) DNA encoding a cytokine;
- b) DNA encoding a mycobacterial heat shock protein gene promoter and translational start site;
- and
- c) a mycobacterial secretion signal sequence, wherein the 5' to 3' order of the components is the DNA of (b), the secretion signal sequence of (c) and the DNA of (a), and the expression of the DNA of (a) is under the control of the heat shock protein gene promoter of (b) and the DNA of (a) replaces the heat shock protein coding sequences.
A recombinant BCG with ability to provide an enhanced immunostimulatory effect in a mammal to which it is administered, said recombinant BCG comprising the following components:
- a) DNA encoding a cytokine selected from the group consisting of:
- IL-2, IFN-.gamma. and GM-CSF;
- b) a mycobacterial heat shock protein gene promoter;
- and
- c) a mycobacterial secretion signal sequence, wherein the 5' to 3' order of the components is the promoter of (b), the secretion signal sequence of (c) and the DNA of (a), the expression of the DNA of (a) is under the control of the promoter of (b) and the cytokine is expressed and secreted from the recombinant BCG.
An E. coli-BCG shuttle plasmid which, when expressed in a mycobacterium, results in enhanced immunostimulatory properties of the mycobacterium, in comparison with immunostimulatory properties of a mycobacterium in which the vector is not expressed, the shuttle plasmid comprising:
- a) DNA encoding a cytokine selected from the group consisting of:
- IL-2, INF-.gamma. and GM-CSF;
- b) a mycobacterial heat shock protein gene promoter;
- and
- c) a mycobacterial secretion signal sequence,
- wherein the 5' to 3' order of the components is the promoter of (b), the secretion signal sequence of (c) and the DNA of (a), and the expression of the DNA of (a) is under the control of the promoter of (b).
A recombinant mycobacterium having enhanced immunostimulatory properties in comparison with immunostimulatory properties of wild type mycobacterium, comprising DNA encoding a cytokine, wherein the DNA is expressed under the control of a promoter and the cytokine is secreted from said mycobacterium in a biologically active form.
Dependencies are inferred from the claim text.