Photochemical internalisation increases the cytotoxic effect of the immunotoxin MOC31-gelonin.
Photochemical internalisation (PCI) was recently demonstrated as a unique procedure for site-specific delivery of several types of membrane impermeable macromolecules from endocytotic vesicles to the cytosol (Berg et al., 1999). The technology is based on the cytosolic release of endocytosed macromolecules from endosomes and lysosomes upon exposure of cells to photosensitising compounds, which became localised to these vesicles, and light. In our study the possibility to increase the cytotoxic effect of the immunotoxin MOC31-gelonin by PCI was examined. The type I ribosome-inactivating protein gelonin was covalently linked to the monoclonal IgG1 antibody MOC31, directed against epithelial glycoprotein-2 (EGP-2), an antigen expressed on most carcinoma cells. Five different cell lines, of which 4 expressed EGP-2, were treated with MOC31-gelonin and endosomal and lysosomal localising photosensitisers, followed by exposure to light. Insignificant cytotoxicity of the MOC31-gelonin was observed when the cells were incubated with the immunotoxin alone. However, in combination with endosomal and lysosomal localising photosensitizers, we demonstrate synergistic toxic effect of the MOC31-gelonin conjugate in a light-dependent manner. Our results indicate that PCI is a promising tool for increasing the cytotoxicity of immunotoxins, which is important for further improvement of the PCI concept towards possible use in cancer therapy.
Selbo
P K
PK
Department of Biophysics, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo, Norway. p.k.selbo@labmed.uio.no
Sivam
G
G
Fodstad
O
O
Sandvig
K
K
Berg
K
K
D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism; D000972:Antineoplastic Agents, Phytogenic / Q000008:administration & dosage / Q000494:pharmacology*; D015815:Cell Adhesion Molecules / Q000276:immunology* / Q000378:metabolism; D004705:Endocytosis; D000071858:Epithelial Cell Adhesion Molecule; D006801:Humans; D000922:Immunotoxins / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000627:therapeutic use; D010940:Plant Proteins / Q000008:administration & dosage / Q000494:pharmacology*; D054789:Ribosome Inactivating Proteins, Type 1; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000972:Antineoplastic Agents, Phytogenic; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; D000922:Immunotoxins; D017319:Photosensitizing Agents; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; C025674:GEL protein, Gelonium multiflorum