Method for selective targeting and entry of bacterial toxins to cells
The present invention provides methods and reagents for directing the selective targeting and entry of bacterial toxins to mammalian cells. Methods include the step of contacting a cell with a bacterial toxin or toxic bioactive fragment thereof associated with an antibody or ligand that is specific to a target on the cell, wherein the antibody or ligand selectively binds the target on the cell and the bacterial toxin or fragment thereof is internalized and enters the cell. The invention further encompasses compositions and kits to carry out the methods.
1. A method for selectively targeting a bacterial toxin to a cell, comprising contacting a cell with a toxic bioactive fragment of a bacterial toxin associated with an antibody or ligand that is specific to a target protein on the cell, wherein the toxic bioactive fragment is associated with the antibody or ligand by an avidin/biotin or streptavidin/biotin linkage, wherein the antibody is selected from the group consisting of a low density lipoprotein receptor antibody, a transferrin receptor antibody, and a vesicular dopamine transport antibody, wherein the toxic bioactive fragment is the enzymatic domain of the bacterial toxin and lacks a functional receptor binding domain; whereby the toxic bioactive fragment is internalized and enters the cell upon selective binding of the antibody or ligand to the target protein.
2. The method according to claim 1 , wherein the bacterial toxin is a clostridium neurotoxin.
3. The method according to claim 1 , wherein the bacterial toxin is selected from the group consisting of botulinum neurotoxin A (BoNT/A), botulinum neurotoxin B (BoNT/B), botulinum neurotoxin C (BoNT/C), botulinum neurotoxin D (BoNT/D), botulinum neurotoxin E (BoNT/E), botulinum neurotoxin F (BoNT/F), botulinum neurotoxin G (BoNT/G), tetanus neurotoxin (TeNT), Diphtheria toxin, and Pseudomonas exotoxin.
4. The method according to claim 1 , wherein the target protein is a plasma membrane protein or a protein located at a plasma membrane of the cell.
5. The method according to claim 1 , wherein the target protein is a receptor protein.
6. The method according to claim 1 , wherein the target protein is a protein located at a secretory vesicle of the cell.
7. The method according to claim 1 , wherein the target protein is a component of the cell's complement system and entry is mediated by said complement system.
8. The method according to claim 1 , wherein the cell is an immune cell, a neuronal cell, an exocrine gland cell, or an endocrine gland cell.
9. The method according to claim 1 , wherein the cell is a cancer or tumor cell.
10. The method according to claim 1 , wherein the cell is contained in a subject.